REVIEW ARTICLE S100–annexin complexes – biology of conditional association Naofumi Miwa1, Tatsuya Uebi2,* and Satoru Kawamura2,3 Department of Physiology, School of Medicine, Toho University, Tokyo, Japan Graduate School of Frontier Biosciences, Osaka University, Japan Department of Biology, Graduate School of Science, Osaka University, Japan Keywords annexin; calcium; colocalization; comprehensive interaction; dicalcin; EF-hand; liposome; membrane trafficking; phospholipid; S100 Correspondence S Kawamura, Graduate School of Frontier Biosciences, Osaka University, Yamada-oka 1–3, Suita, Osaka 565-0871, Japan Fax: +81 6879 4614 Tel: +81 6879 4610 E-mail: kawamura@fbs.osaka-u.ac.jp *Present address Laboratory of Cell Signal and Metabolism, National Institute of Biomedical Innovation, Osaka, Japan (Received 17 June 2008, revised August 2008, accepted 22 August 2008) doi:10.1111/j.1742-4658.2008.06653.x S100 proteins and annexins both constitute groups of Ca2+-binding proteins, each of which comprises more than 10 members S100 proteins are small, dimeric, EF-hand-type Ca2+-binding proteins that exert both intracellular and extracellular functions Within the cells, S100 proteins regulate various reactions, including phosphorylation, in response to changes in the intracellular Ca2+ concentration Although S100 proteins are known to be associated with many diseases, exact pathological contributions have not been proven in detail Annexins are non-EF-hand-type Ca2+-binding proteins that exhibit Ca2+-dependent binding to phospholipids and membranes in various tissues Annexins bring different membranes into proximity and assist them to fuse, and therefore are believed to play a role in membrane trafficking and organization Several S100 proteins and annexins are known to interact with each other in either a Ca2+-dependent or Ca2+-independent manner, and form complexes that exhibit biological activities This review focuses on the interaction between S100 proteins and annexins, and the possible biological roles of these complexes Recent studies have shown that S100–annexin complexes have a role in the differentiation of gonad cells and neurological disorders, such as depression These complexes regulate the organization of membranes and vesicles, and thereby may participate in the appropriate disposition of membrane-associated proteins, including ion channels and ⁄ or receptors Introduction The interaction between S100 and annexin proteins was initially identified in porcine intestinal brush border-derived membranes, as a complex formed between S100A10 and annexin A2 Annexin A2 (previously named p36 or calpactin I, etc.) is a substrate of srcrelated viral tyrosine kinase [1,2], which raises the possibility that this complex may be involved in cancer-related pathology The complex of S100A10 and annexin A2 (S100A10–annexin A2 complex) has been found to bind to cytoskeletal components and to colocalize in submembranous compartments [3], suggesting that this complex may play a role in subcellular vesicle organization to exert its biological function Following these findings, another S100 member, S100A11 (originally named S100C or calgizzarin), was found to interact with annexin A1 in a Ca2+-dependent manner, with additional evidence showing that this complex also binds to cytoskeletal components, such as tubulin and vimentin Unlike the interaction between S100A10 and annexin A2, the interaction between S100A11 and annexin A1 occurs in a temporal FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4945 Biology of S100–annexin complexes N Miwa et al manner when the intracellular Ca2+ level increases, and therefore this complex has been postulated to regulate Ca2+-dependent membrane organization during vesiculation or internalization To date, several other pairs of S100 proteins and annexins have been reported (Table 1), and it seems timely to view these pairs as constituents of a broad system of S100–annexin complexes In this system, some S100 proteins are able to bind to several annexins The host (for example, S100 protein) and its binding partner (an annexin protein) can be determined by their subcellular distributions and temporal expression patterns in each tissue In this review, after a brief description of S100 proteins, annexins and our recently characterized dicalcin, an S100-like protein, we review several well-characterized S100–annexin complexes to obtain an understanding of the divergence of the physiological roles of the different complexes The structural basis of complex formation is reviewed in the accompanying article by RintalaDempsey et al [4] Proteins sues, including brain, lung and heart An important feature of S100 proteins is their role as Ca2+ sensors Each S100 protein has a pair of high-affinity Ca2+binding sites, called EF-hand motifs When intracellular Ca2+ concentrations increase after environmental stimuli, for example, S100 proteins can bind to Ca2+ via EF-hand motifs and undergo large conformational changes These changes induce the exposure of a hydrophobic patch at the surface of these molecules and assist them to interact with their target proteins, including enzymes (e.g kinase, phospholipase A2) and cytoskeletal proteins (e.g actin) In this way, S100 proteins transduce environmental signals to intracellular activities to regulate cell proliferation, differentiation, etc [7,8] Some S100 members are secreted from cells through undefined exocytotic machinery, exerting extracellular actions, such as anti-apoptosis and anti-coagulation, through their receptors on the surface of the plasma membrane A number of targets have been reported to date [9], and, for several S100 members, genetically engineered animals have been produced to study the functional role(s) of S100 proteins [10] S100 proteins Annexins S100 proteins form a family of small (10–14 kDa) Ca2+-binding proteins that regulate various intracellular and extracellular processes Increased levels of S100 proteins have been reported to be associated with a number of diseases Originally, S100A1 (originally named S100a) and S100B (S100b) were isolated in bovine brain as proteins soluble in 100% (saturated) ammonium sulfate at neutral pH [5] To date, 20 S100 genes have been identified exclusively in vertebrates, including humans, with most of the S100 genes clustered on human chromosome 1q21 (S100A1– S100A16), whereas no S100 genes have been detected in invertebrates [6] S100 proteins are known to exist as homo- ⁄ heterodimeric functional units in various tis- Annexins are another family of Ca2+-binding proteins Their Ca2+-binding motifs are different from the EF-hand type described above and are called annexin type or type II [11,12] On Ca2+ binding, annexins can interact with anionic membrane phospholipids, making them ‘Ca2+-dependent phospholipid-binding proteins’ Annexins were first identified from several sources and were given different names (e.g lipocortin, calpactin, etc.) Later, these proteins were given a new family name of ‘annexin’, because the major property of this family is to ‘annex’ cellular membranes in a Ca2+-dependent manner [13] Annexins are distributed in various species from humans to plants, and, to date, the vertebrate annex- Table Complex formation between S100 proteins and annexins An S100–annexin complex is formed as indicated by the reference numbers Annexin A1 S100A1 S100A4 S100A6 S100A10 S100A11 S100A12 S100B 4946 Annexin A2 [47,51,52,57–59,63] Annexin A5 [61] [60] [3,22–46] [58] Annexin A6 Annexin A11 [82,83] [88] [66,70,71,74] [62] [89] [82,83] FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS N Miwa et al ins, which have been most extensively studied, comprise up to 12 members [14] Annexins are expressed widely in many tissues, but their localization varies: some are present intracellularly and others are localized at the plasma membrane Most annexins consist of an individually unique N-terminal domain and a fairly conserved C-terminal core that contains either four or eight repeating units of approximately 70 amino acids It is believed that the annexin C-terminal core is a module that mediates both Ca2+ and membrane binding Annexins interact with many targets and exert various biological functions, including regulation of membrane aggregation and membrane trafficking They also have extracellular functions, for example, in anti-inflammation and anti-coagulation [11,12] Although a few annexins have been analysed in knockout animals [14,15], their phenotypes are subtle, so that their exact physiological functions remain elusive Dicalcin Dicalcin, an S100-like Ca2+-binding protein formerly called p26olf, was originally identified in frog (Rana catesbeiana) olfactory epithelium [16] After the original identification, however, this protein was also found in other tissues, including lung and spleen Although detailed structural analysis (i.e crystallographic study) has not been carried out, sequence alignment and molecular modelling have suggested that dicalcin consists of two S100-like regions aligned in tandem (each region has approximately 50% identity to the sequence of chick S100A11), and possibly adopts a remarkably similar conformation to that of a homodimeric form of S100B [17,18] As all other S100 members, except calbindin, form a homo- or heterodimer in solution to exert their biological functions, dicalcin may substitute the function(s) of S100 proteins in the form of a monomer Based on this consideration, we gave it a mnemonic name: ‘dimer form of S100 calcium-binding protein’ Our quantitative Ca2+-binding study showed cooperative Ca2+ binding of dicalcin, with an apparent overall dissociation constant (Kd) of 10–20 lm [19] On Ca2+ binding, dicalcin interacts with a set of annexin members in both the olfactory and respiratory cilia [20], as well as with several other olfactory cilia proteins, including b-adrenergic receptor-like protein, which has not yet been cloned [21] Through interactions with annexins, dicalcin enhances liposome aggregation in a Ca2+-dependent manner, which suggests that dicalcin plays a role in membrane-associated events in the olfactory and respiratory cilia (see below) Biology of S100–annexin complexes S100–annexin complexes S100A10–annexin A2 complex Distribution The mRNA expression of S100A10 and annexin A2 has been shown in various mouse tissues, and both are expressed coincidentally at high levels in lung, intestine and thymus [22] On the basis of an immunohistochemical colocalization study [3], both S100A10 and annexin A2 were found in the following sites: (a) brush border in porcine intestine; (b) glomerular cells including mesangial cells and endothelial cells in porcine kidney; (c) endothelial cells in porcine brain; and (d) fibroblasts in bovine heart Within these cells, both proteins were mainly localized to endosomes and at the plasma membrane [23–25] Properties of interaction S100A10 (alternatively called p11) and annexin A2 are known to exist as a heterotetramer [(S100A10)2–(annexin A2)2] in a membrane fraction [26] The S100A10binding site in annexin A2 is considered to reside in N-terminal residues (Val3, Ile6, Leu7, Leu10) based on cosedimentation and gel filtration experiments using truncated annexin mutants [27,28] S100A10 is an exceptional protein amongst S100 members in terms of Ca2+ binding: S100A10 is unable to bind to Ca2+ because of a mutation within its EF-hand motifs Three amino acid residues are lost in the N-terminal EF-hand motif and crucial amino acids are substituted in the C-terminal motif [29] As a consequence, the association of S100A10 and annexin A2 is Ca2+ independent: these two proteins form a heterotetrameric complex constitutively regardless of the Ca2+ concentration Instead of Ca2+, post-translational modifications of annexin A2 have regulatory effects on the association with S100A10: N-acetylation of annexin A2 is necessary for this association [30,31] and protein kinase C-mediated phosphorylation decreases the affinity of annexin A2 for S100A10 [32] Binding targets of the complex In an S100A10–annexin A2 complex, an S100A10 dimer resides in the centre of the complex, interconnecting two annexin A2 molecules [26] Annexins in the outer position of this complex preferentially bind to anionic phospholipids, such as phosphatidylinositol 4,5-bisphosphate, which is enriched in lipid rafts in the plasma membrane Because S100A10 has the ability to bind to cytoskeletal proteins, such as actin, this FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4947 Biology of S100–annexin complexes N Miwa et al complex can link membranes and ⁄ or vesicles to cytoskeletal proteins to regulate membrane organization This association of an S100–annexin A2 complex with lipid membranes is Ca2+ dependent with a Kd value of lm [33], which probably reflects Ca2+ binding to annexin A2 (S100A10 does not have Ca2+-binding ability) The S100–annexin A2 complex has also been shown to interact with membrane-related proteins They include certain types of sodium channel [34], potassium channels [35,36], transient receptor potential channels [37] and serotonin 5-HT1B receptors [38] The molecular topology of this complex in the membrane-bound state has been postulated from two scenarios derived from different experimental approaches Cryoelectron microscopy has suggested that each annexin A2 molecule in the outer position of the complex binds to one membrane, and therefore the tetrameric complex links two different membranes [39] By contrast, scanning force microscopy has suggested that two annexin A2 molecules bind to the same membrane [40] In the latter case, the S100A10 dimer resides in a relatively outer position of the complex away from the membrane, and thereby interacts with other proteins (e.g cytosolic portion of channels or receptors), enabling them to be associated with or incorporated into the membranes that are bound by annexin A2 molecules In addition to the intracellular targets described above, the S100A10–annexin A2 complex has been shown to bind to tissue-type plasminogen activator in the extracellular space and to act as a functional receptor to produce plasminogen from tissue-type plasminogen activator [41] However, the exact binding character remains a matter of debate [42] Biological roles Several studies using knockout animals have suggested the biological roles of this complex [43] Foulkes et al [44] have demonstrated that S100A10– ⁄ – mice show deficient nociception, which may be attributed to a severe decrease in the sodium current Svenningsson et al [38] have found that S100A10) ⁄ ) mice exhibit a depression-like phenotype with reduced responses to 5-HT1B agonists; this suggests that the lack of this complex causes a depressive disorder Recently, this group has also shown that S100A10 has an inhibitory role on some abnormal behaviors caused by l-3,4-dihydroxyphenylalanine administration to an animal model of Parkinsonism [45] The identification of the targets of the S100A10–annexin A2 complex (see above) led to the suggestion that this complex functions as a guiding molecule of channels and ⁄ or 4948 receptors from the endoplasmic reticulum to the Golgi and ⁄ or internalized vesicle to the plasma membrane The deficits in these knockout animals may be attributed to the improper association with or incorporation into the plasma membrane of these channels and ⁄ or receptors Another possible biological role of this complex is related to fibrin homeostasis In the normal blood vessel, fibrin is not deposited and arterial thrombin is cleared after injury However, S100A10) ⁄ ) knockout animals show a displaced deposition of fibrin in the microvasculature and incomplete clearance of arterial thrombin [46]; this may be caused by the loss of the S100A10–annexin A2 complex on the outer surface of the plasma membrane of the endothelial cells S100A11–annexin A1 complex Distribution The association of S100A11 (previously known as S100C or calgizzarin) with annexin A1 was initially found during the search for targets of annexin A1, a prototype of annexin that has attracted considerable interest because of its involvement in cell growth and differentiation [47] S100A11 mRNA is distributed in almost all human tissues It is highly expressed in muscle, heart and bladder [48,49] Annexin A1 is also widely expressed in many tissues, including lung, kidney and spleen [50] Within the cells, annexin A1 is localized mostly in the cytosol, except for its presence within nuclei of the human respiratory epithelium [50] Although the subcellular colocalization of these two proteins in vivo has not been studied in detail, ectopically expressed S100A11 has been shown to colocalize with intrinsic annexin A1 on the early endosomal membranes of fibroblastic BHK cells [51] Biochemical studies have shown that S100A11 and annexin A1 are both present in the cornified envelope preparation of human keratinocytes [52] Properties of interaction In contrast with the interaction between S100A10 and annexin A2, S100A11 binds to annexin A1 in a Ca2+dependent manner [47], evoking the suggestion that this complex regulates Ca2+-dependent cellular events S100A11 has been shown to bind to annexin A1 at high Ca2+ concentrations (1 mm), presumably forming a heterotetramer [(S100A11)2–(annexin A1)2] [47] As an individual protein, S100A11 alone binds to Ca2+ with a Kd value of 8–16 lm [53] and undergoes conformational changes with a half-maximal effective Ca2+ concentration at a similar concentration FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS N Miwa et al ( 35 lm) in measurements with fluorescent-labelled probes [54] Annexin A1 alone binds to Ca2+ with a Kd value of 20–75 lm, enhancing its binding activity for phospholipid vesicles [55,56] Although detailed analysis of the Ca2+ concentration required for the association of S100A11 with annexin A1 has not yet been carried out, these two proteins have been hypothesized to associate within a similar Ca2+ concentration range in which both S100A11 and annexin A1 can bind to Ca2+ The S100A11-binding site in annexin A1 is considered to reside in the N-terminal residues, as revealed by experiments similar to those used for the identification of the S100A10-binding site in annexin A2 [47,57,58] With regard to the specificity of S100A11 binding to annexin members, a previous study using fluorescent-labelled peptides has shown that S100A11 interacts specifically with the annexin A1 N-terminal domain and does not interact with the corresponding N-terminal domain of annexin A2 [59] However, a recent study using annexin A2 peptides has shown that S100A11 also interacts with the N-terminal domain of annexin A2 [58], consistent with the finding that annexin A2 shows broad binding specificity to other S100 members (e.g S100A4 and S100A6) [60,61] Binding of S100A11 to both annexins A1 and A2 suggests possible multifunctional roles of S100A11 in the regulation of membrane trafficking and ⁄ or organization Binding targets and roles of the complex In contrast with the detailed structural analysis of the S100A11–annexin A1 complex, the cellular targets and functions of this complex have not been studied in detail Potential targets of this complex may be phospholipids and cytoskeletal proteins based on the consideration of the following reports: (a) annexin A1 alone binds to lipid membranes in a Ca2+-dependent manner [55,56]; (b) S100A11 alone binds to cytoskeletal proteins with a Kd value of lm in porcine heart [53]; (c) S100A11 is also able to interact with annexin A6 at a high Ca2+ concentration (1 mm), and this S100A11–annexin A6 complex binds to native liposomes derived from rat vascular smooth muscle as well as phosphatidylserine liposomes in the presence of Ca2+ (200 lm) [62] With regard to a potential biological role(s) of the S100A11–annexin A1 complex, Robinson et al [52] have reported that S100A11 and annexin A1 are colocalized beneath the plasma membrane during the final stages of epidermal keratinocyte differentiation, indicating that this complex may be involved in the formation of the cornified envelope in human keratinocytes Biology of S100–annexin complexes A biochemical study has shown that S100A11 suppresses the phosphorylation of annexin A1 by protein kinase C, resulting in a decrease in the aggregation of phospholipid vesicles [63] This result also suggests a role for the S100A11–annexin A1 complex in the regulation of membrane organization S100A11 has been shown to inhibit actin-activated myosin Mg2+-ATPase activity in a Ca2+-dependent manner and to regulate the generation of smooth muscle force with a Kd value of 50 lm [64] In smooth muscle, however, annexin A1 is not expressed abundantly [50], and therefore the S100A11–annexin A1 complex may not be involved in this biological effect S100A6–annexin A11 complex Distribution Both S100A6 (formally called calcyclin) and annexin A11 have been studied to investigate their involvement in cell cycle regulation and cancer biology, because the expression levels of these proteins are high in malignant tumours [65,66] S100A6 is expressed in smooth muscle cells, epithelial cells and fibroblasts in almost all mammalian tissues, including intestine, kidney [67,68] and brain [69] Within these cells, S100A6 is expressed at the plasma membrane and the nuclear envelope in embryonic pig testis-derived ST cell lines, as well as human skin and embryonic mouse testis [66,70,71] The expression level of S100A6 is elevated in a number of malignant tumours, such as acute myeloid leukaemia, neuroblastoma and melanoma cell lines [72,73], with peak expression between the G0 and S phases of the cell cycle [68,74,75] Annexin A11 is also widely distributed in the nucleoplasm in many cultured cell lines The subcellular distribution of annexin A11 is altered during the cell cycle: it shows a dynamic and biphasic interaction with the nuclear envelope, first during envelope breakdown and second during its reassembly [66] Properties of interaction and targets of the complex Ca2+-dependent interaction of S100A6 and annexin A11 was initially found in biochemical S100A6 affinity chromatography [76] However, our knowledge of this interaction (e.g binding property and molecular target of the complex) is still limited S100A6 has been shown to bind to the N-terminus (Gln49–Thr62) of annexin A11 at approximately 200 lm Ca2+ This S100A6–annexin A11 complex has been shown to bind to phospholipid vesicles in the presence of Ca2+ (1 mm) [76] FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4949 Biology of S100–annexin complexes N Miwa et al Biological roles Biological roles of the complexes S100A6 was originally identified as a cDNA clone for which cognate RNA was growth regulated [65], and subsequently purified as a protein [77,78] S100A6 has been shown to interact with the nuclear envelope in a Ca2+-dependent manner, as does annexin A11, and subsequently both were found to be colocalized in proliferating cells during certain stages in the cell cycle [66,70] In epidermoid carcinoma A431 cells and vascular smooth muscle cells, an increase in the Ca2+ concentration, especially during the prophase, leads to the translocation of annexin A11 from the nucleus to the nuclear envelope, where it is colocalized with S100A6 [66], suggesting a role of this complex in cell cycle regulation In addition, S100A6 and annexin A11 have been shown to be colocalized in mouse gonad during an important period for male–female determination, suggesting that this complex plays a role in cell stage-specific events that trigger a cascade for sex determination [71] Both S100A1 and S100B alone have been shown to hamper the assembly of glial fibrillary acidic protein and desmin, and to inhibit the formation of intermediate filaments in vitro [85,86] However, this inhibitory effect was lost when the C-terminal core, but not the N-terminal domain, of annexin A6 was added [83] The molecular mechanism of this effect is, however, unknown, and therefore it is not certain whether this effect is brought about by a ‘passive’ decrease in the amount of effective S100 protein as a result of its adsorption to annexin A6, or by an ‘active’ action mediated by a target molecule(s) of the complex Alternatively, these complexes have been suggested to play a role in the regulation of Ca2+ fluxes in skeletal muscle cells by affecting a ryanodine receptor in the sarcoplasmic reticulum [82] S100A1–annexin A6 and S100B–annexin A6 complexes Distribution S100A1 is expressed in a variety of tissues, including the nervous system, skeletal muscle, heart, kidney and fat [79] S100B is abundant in the nervous system, testis, fat, skin and cartilage [80] Annexin A6 is expressed as two isoforms, a long form (annexin A6-1) and a short form (annexin A6-2), determined by alternative splicing [81] Both isoforms prevail in a variety of tissues, including kidney, heart and skeletal muscle, with predominant expression of annexin A6-1 [81] S100A1, S100B and annexin A6 have been shown to colocalize in the sarcolemma, the membranes of the sarcoplasmic reticulum and transverse tubules in avian skeletal muscle cells [82] Properties of interaction and targets of the complex A biochemical study using fluorescent-labelled proteins has shown that both S100A1 and S100B interact with annexin A6 at high Ca2+ concentrations (100 lm) [83], and both the N-terminal domain and the C-terminal core of annexin A6 bind to S100 proteins The target molecules and cellular structures of these two complexes have not been identified Although several combinations of S100 proteins and annexins are known to bind to liposomes (see above), the S100A1–annexin A6 and S100B– annexin A6 complexes showed no apparent interactions with liposomes in a cosedimentation assay [84] 4950 Dicalcin–annexin complex Distribution Dicalcin is expressed in a variety of frog tissues [16] In the olfactory and respiratory epithelium, dicalcin and annexins A1, A2 and A5 are all localized in the cilia of these tissues [20]; furthermore, all four proteins are colocalized in the same cilia Western analysis using a Chaps-solubilized cilia membrane fraction indicated that the ratio of the content of annexins and dicalcin were A1 : A2 : A5 : dicalcin = :  0.42 :  0.54 :  1.9, and this estimated content of dicalcin seems to be sufficient to interact with all members of annexins expressed in the cilia [20] Properties of interaction and targets of the complex Dicalcin and annexins (annexins A1, A2 and A5) form a complex in a Ca2+-dependent manner, as revealed by Ca2+-dependent binding of annexins to dicalcinconjugated Sepharose Although other S100 members have been shown to bind to the N-terminus of annexins (see above), dicalcin binds to N-terminal truncated annexins, indicating that the C-terminal core alone is capable of binding to dicalcin [20] Indeed, each of the frog annexins A1, A2 and A5 has at least a few putative S100-binding motifs in the C-terminal core: for example, in annexin A2, the consensus sequence FXFFXXF (where F denotes a hydrophobic residue and X is any amino acid; [62]) can be found in L54– V60 and L257–I263 [20] However, a recent study has shown that full-length annexin A2 possesses an approximately four- to five-fold increased capacity for binding to dicalcin-conjugated Sepharose, relative to that of FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS N Miwa et al N-terminal truncated annexin A2 (T Uebi, N Miwa and S Kawamura, unpublished results), indicating the involvement of the N-terminus of annexin A2 in its binding to dicalcin The binding affinity of the N-terminus or the core domain has not yet been determined The binding of dicalcin–annexins to liposomes has been examined As shown above, annexins A1 and A2, by themselves, exhibit activities to induce liposome aggregation in a Ca2+-dependent manner Remarkably, dicalcin enhances this liposome aggregation activity of annexin A1 and A2, but shows little effect on the activity of annexin A5 [20] As our assay mixture contained only dicalcin, annexins and liposomes, the dicalcin–annexin A1 and dicalcin–annexin A2 complexes are likely to bind directly to liposomes and to enhance liposome aggregation The effective Ca2+ concentration for liposome aggregation depends on which annexin binds to dicalcin Half-maximal effects with dicalcin–annexin A1 and dicalcin–annexin A2 complexes were observed at approximately 30 lm and < lm Ca2+, respectively These effective Ca2+ concentrations did not change significantly in the presence or absence of dicalcin, and therefore the difference in the Ca2+ concentration for half-maximal effects between the two complexes can be attributed to the different affinity of each annexin for Ca2+ As described above, dicalcin probably binds to two molecules of annexin To determine whether dicalcin binds to two of the same subtype of annexin or to two different subtypes, we measured Ca2+- and dicalcindependent liposome aggregation in the presence of a mixture of annexins of different subtypes The profile of liposome aggregation was simply the sum of the results obtained with a single subtype of annexin, suggesting that dicalcin tends to bind to the same subtype of annexin, even in the presence of different subtypes in a mixture Biological roles of the complexes Dicalcin and annexins are colocalized in olfactory and respiratory cilia which are motile Motile cells are often subject to mechanical stress and damage [87] In addition, olfactory cilia are exposed to environmental chemicals, microorganisms and viruses, so that the cilia membrane is often likely to be damaged and disrupted Therefore, the cytoplasmic Ca2+ concentration at the disrupted site may increase in a variable manner according to the severity of the damage, and sometimes increase even to the extracellular level (a few mm) Dicalcin–annexin complexes are able to regulate membrane aggregation within a wide range of Ca2+ concentration by utilizing two annexin subtypes that Biology of S100–annexin complexes cover different Ca2+ concentrations This mechanism may serve to reseal the cilia membrane in response to a wide range of Ca2+ increases caused by disruption of these membranes [20] In this sense, dicalcin– annexin complexes in the olfactory and respiratory cilia may be a typical example of a system in which different subtypes of family proteins act in a complementary manner to cover a wide range of changes in intracellular conditions In addition to annexins, dicalcin has been shown to interact with several olfactory cilia proteins in a Ca2+dependent manner [21] One possible candidate is olfactory b-adrenergic receptor kinase-like protein Considering the possible role of annexins in membrane organization, we hypothesize that the dicalcin–annexin complex could bind to a protein, such as b-adrenergic receptor kinase-like protein, to incorporate or associate the protein into the membranes, as is postulated for the S100A10–annexin A2 complex (see above) Other S100–annexin complexes Although the number of reports is limited, other S100– annexin complexes have been reported: S100A4–annexin A2 [61], S100A6–annexin A2 [60], S100A6–annexin A6 [88], S100A11–annexin A2 [58], S100A11–annexin A6 [62] and S100A12–annexin A5 [89] (see Table 1) Future perspectives As discussed above, various pairing of S100 and annexins may be an intrinsic and conventional mechanism of the S-100 annexin system to function in a variety of tissues The participants of these complexes are likely to be determined by their spatial and temporal distribution patterns in cells By switching partners, an S100–annexin complex may exhibit tissue- and cell stage-specific biological actions, such as the regulation of cell cycle and membrane traffic Our current knowledge of this system is still fragmentary, and the exact molecular mechanisms remain unknown For a better understanding of the S100–annexin system, further investigations are certainly required As shown in this review, some S100–annexin pairs exhibit broad binding specificity These proteins may interact with a less favourable member protein in the absence of their most favourable partner, and this complex may possibly substitute for the function of the complex of the most favourable pair This may be the reason why only subtle changes are observed in the phenotype of knockout animals of S100 proteins and annexins Therefore, there is a need to generate multiple knockout animals deficient in several S100 and ⁄ or annexin FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4951 Biology of S100–annexin complexes N Miwa et al proteins in order to reveal distinctive phenotypic changes 17 References Gerke V & Weber K (1984) Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin EMBO J 3, 227–233 Glenney JR Jr (1985) Phosphorylation of p36 in vitro with pp60src Regulation by Ca2+ and phospholipid FEBS Lett 192, 79–82 Osborn M, Johnsson N, Wehland J & Weber K (1988) The submembranous location of p11 and its interaction with the p36 substrate of pp60 src kinase in situ Exp Cell Res 175, 81–96 Rintala-Dempsey AC, Rezvanpour A & Shaw GS (2008) S100–annexin complexes – structural insights Febs J 275, 4945–4955 Moore BW (1965) A soluble protein characteristic of the nervous system Biochem Biophys Res Commun 19, 739–744 Marenholz I, Heizmann CW & Fritz G (2004) S100 proteins in mouse and man: from evolution to function and pathology (including an update of the nomenclature) Biochem Biophys Res Commun 322, 1111–1122 Heizmann CW, Fritz G & Schafer BW (2002) S100 proă teins: structure, functions and pathology Front Biosci 7, 1356–1368 Donato R (2003) Intracellular and extracellular roles of S100 proteins Microsci Res Tech 60, 540–551 Santamaria-Kisiel L, Rintala-Dempsey AC & Shaw GS (2006) Calcium-dependent and -independent interactions of the S100 protein family Biochem J 396, 201–214 10 Heizmann CW, Ackermann GE & Galichet A (2007) Pathologies involving the S100 proteins and RAGE Subcell Biochem 45, 93–138 11 Raynal P & Pollard HB (1994) Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins Biochim Biophys Acta 1197, 63–93 12 Gerke V & Moss SE (2002) Annexins: from structure to function Physiol Rev 82, 331–371 13 Crumpton MJ & Dedman JR (1990) Protein terminology tangle Nature 345, 212 14 Rescher U & Gerke V (2004) Annexins – unique membrane binding proteins with diverse functions J Cell Sci 117, 2631–2639 15 Hayes MJ & Moss SE (2004) Annexins and disease Biochem Biophys Res Commun 322, 1166–1170 16 Miwa N, Kobayashi M, Takamatsu K & Kawamura S (1998) Purification and molecular cloning of a novel calcium-binding protein, p26olf, in the frog olfactory 4952 18 19 20 21 22 23 24 25 26 27 28 29 epithelium Biochem Biophys Res Commun 251, 860– 867 Tanaka T, Miwa N, Kawamura S, Sohma H, Nitta K & Matsushima N (1999) Molecular modeling of single polypeptide chain of calcium-binding protein p26olf from dimeric S100B(bb) Protein Eng 12, 395–405 Miwa N & Kawamura S (2003) Frog p26olf, a molecule with two S100-like regions in a single peptide Microsci Res Tech 60, 593–599 Miwa N, Shinmyo Y & Kawamura S (2001) Calciumbinding by p26olf, an S100-like protein in the frog olfactory epithelium Eur J Biochem 268, 6029–6036 Uebi T, Miwa N & Kawamura S (2007) Comprehensive interaction of dicalcin with annexins in frog olfactory and respiratory cilia FEBS J 274, 4863–4876 Miwa N, Uebi T & Kawamura S (2000) Characterization of p26olf, a novel calcium-binding protein in the frog olfactory epithelium J Biol Chem 275, 27245– 27249 Saris CJ, Kristensen T, D’Eustachio P, Hicks LJ, Noonan DJ, Hunter T & Tack BF (1987) cDNA sequence and tissue distribution of the mRNA for bovine and murine p11, the S100-related light chain of the proteintyrosine kinase substrate p36 (calpactin I) J Biol Chem 262, 10663–10671 Zokas L & Glenney JR Jr (1987) The calpactin light chain is tightly linked to the cytoskeletal form of calpactin I: studies using monoclonal antibodies to calpactin subunits J Cell Biol 105, 2111–2121 Thiel C, Osborn M & Gerke V (1992) The tight association of the tyrosine kinase substrate annexin II with the submembranous cytoskeleton depends on intact p11and Ca2+-binding sites J Cell Sci 103, 733–742 Chasserot-Golaz S, Vitale N, Sagot I, Delouche B, Dirrig S, Pradel LA, Henry JP, Aunis D & Bader MF (1996) Annexin II in exocytosis: catecholamine secretion requires the translocation of p36 to the subplasmalemmal region in chromaffin cells J Cell Biol 133, 1217– 1236 Sopkova-de Oliveira Santos J, OLing FK, Rety S, Brisson A, Smith JC & Lewit-Bentley A (2000) S110 protein–annexin interactions: a model of the (Anx2– p11)2 heterotetramer complex Biochim Biophys Acta 1498, 181–191 Glenney JR Jr, Boudreau M, Galyean R, Hunter T & Tack B (1986) Association of the S-100-related calpactin I light chain with the NH2-terminal tail of the 36-kDa heavy chain J Biol Chem 261, 10485–10488 Kube E, Becker T, Weber K & Gerke V (1992) Protein–protein interaction studied by site-directed mutagenesis Characterization of the annexin II-binding site on p11, a member of the S100 protein family J Biol Chem 267, 14175–14182 Gerke V & Weber K (1985) Calcium-dependent conformational changes in the 36-kDa subunit of intestinal FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS N Miwa et al 30 31 32 33 34 35 36 37 38 39 40 41 protein I related to the cellular 36-kDa target of Rous sarcoma virus tyrosine kinase J Biol Chem 260, 1688– 1695 Johnsson N, Marriott G & Weber K (1988) p36, the major cytoplasmic substrate of src tyrosine protein kinase, binds to its p11 regulatory subunit via a short amino-terminal amphipathic helix EMBO J 7, 2435– 2442 Becker T, Weber K & Johnsson N (1990) Protein–protein recognition via short amphiphilic helices; a mutational analysis of the binding site of annexin II for p11 EMBO J 9, 4207–4213 Jost M & Gerke V (1996) Mapping of a regulatory important site for protein kinase C phosphorylation in the N-terminal domain of annexin II Biochim Biophys Acta 1313, 283–289 Drust DS & Creutz CE (1988) Aggregation of chromaffin granules by calpactin at micromolar levels of calcium Nature 331, 88–91 Okuse K, Malik-Hall M, Baker MD, Poon WY, Kong H, Chao MV & Wood JN (2002) Annexin II light chain regulates sensory neuron-specific sodium channel expression Nature 417, 653–656 Girard C, Tinel N, Terrenoire C, Romey G, Lazdunski M & Borsotto M (2002) p11, an annexin II subunit, an auxiliary protein associated with the background K+ channel, TASK-1 EMBO J 21, 4439–4448 ´ Renigunta V, Yuan H, Zuzarte M, Rinne S, Koch A, Wischmeyer E, Schlichthorl G, Gao Y, Karschin A, ¨ Jacob R et al (2006) The retention factor p11 confers an endoplasmic reticulum-localization signal to the potassium channel TASK-1 Traffic 7, 168– 181 van de Graaf SF, Hoenderop JG, Gkika D, Lamers D, Prenen J, Rescher U, Gerke V, Staub O, Nilius B & Bindels RJ (2003) Functional expression of the epithelial Ca2+ channels (TRPV5 and TRPV6) requires association of the S100A10–annexin complex EMBO J 22, 1478–1487 Svenningsson P, Chergui K, Rachleff I, Flajolet M, Zhang X, El Yacoubi M, Vaugeois JM, Nomikos GG & Greengard P (2006) Alterations in 5-HT1B receptor function by p11 in depression-like states Science 311, 77–80 Lambert O, Gerke V, Bader MF, Porte F & Brisson A (1997) Structural analysis of junctions formed between lipid membranes and several annexins by cryo-electron microscopy J Mol Biol 272, 42–55 Menke M, Ross M, Gerke V & Steinem C (2004) The molecular arrangement of membrane-bound annexin A2–S100A10 tetramer as revealed by scanning force microscopy Chembiochem 5, 1003–1006 Cesarman-Maus G & Hajjar KA (2005) Molecular mechanisms of fibrinolysis Br J Haematol 129, 307– 321 Biology of S100–annexin complexes 42 Waisman DM (2005) Annexin A2 may not play a role as a plasminogen receptor Br J Haematol 131, 552– 556 43 Rescher U & Gerke V (2008) S100A10 ⁄ p11: family, friends and functions Pflugers Arch 455, 575–582 44 Foulkes T, Nassar MA, Lane T, Matthews EA, Baker MD, Gerke V, Okuse K, Dickenson AH & Wood JN (2006) Deletion of annexin light chain p11 in nociceptors causes deficits in somatosensory coding and pain behavior J Neurosci 26, 10499–10507 45 Zhang X, Andren PE, Greengard P & Svenningsson P (2008) Evidence for a role of the 5-HT1B receptor and its adaptor protein, p11, in L-DOPA treatment of an animal model of Parkinsonism Proc Natl Acad Sci USA 105, 2163–2168 46 Ling Q, Jacovina AT, Deora A, Febbraio M, Simantov R, Silverstein RL, Hempstead B, Mark WH & Hajjar KA (2004) Annexin II regulates fibrin homeostasis and neoangiogenesis in vivo J Clin Invest 113, 38–48 47 Mailliard WS, Haigler HT & Schlaepfer DD (1996) Calcium-dependent binding of S100C to the N-terminal domain of annexin I J Biol Chem 271, 719–725 48 Ohta H, Sasaki T, Naka M, Hiraoka O, Miyamoto C, Furuichi Y & Tanaka T (1991) Molecular cloning and expression of the cDNA coding for a new member of the S100 protein family from porcine cardiac muscle FEBS Lett 295, 93–96 49 Inada H, Naka M, Tanaka T, Davey GE & Heizmann CW (1999) Human S100A11 exhibits differential steady-state RNA levels in various tissues and a distinct subcellular localization Biochem Biophys Res Commun 263, 135–138 50 Dreier R, Schmid KW & Gerke V (1998) Differential expression of annexins I, II and IV in human tissues: an immunohistochemical study Histochem Cell Biol 110, 137–148 51 Seemann J, Weber K & Gerke V (1997) Annexin I targets S100C to early endosomes FEBS Lett 413, 185–190 52 Robinson NA, Lapic S, Welter JF & Eckert RL (1997) S100A11, S100A10, annexin I, desmosomal proteins, small proline-rich proteins, plasminogen activator inhibitor-2, and involucrin are components of the cornified envelope of cultured human epidermal keratinocytes J Biol Chem 272, 12035–12046 53 Naka M, Qing ZX, Sasaki T, Kise H, Tawara I, Hamaguchi S & Tanaka T (1994) Purification and characterization of a novel calcium-binding protein, S100C, from porcine heart Biochim Biophys Acta 1223, 348– 353 54 Allen BG, Durussel I, Walsh MP & Cox JA (1996) Characterization of the Ca2+-binding properties of calgizzarin (S100C) isolated from chicken gizzard smooth muscle Biochem Cell Biol 74, 687–694 55 Schlaepfer DD & Haigler HT (1987) Characterization of Ca2+-dependent phospholipid binding and FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4953 Biology of S100–annexin complexes 56 57 58 59 60 61 62 63 64 65 66 67 N Miwa et al phosphorylation of lipocortin I J Biol Chem 262, 6931– 6937 Ando Y, Imamura S, Hong YM, Owada MK, Kakunaga T & Kannagi R (1989) Enhancement of calcium sensitivity of lipocortin I in phospholipid binding induced by limited proteolysis and phosphorylation at the amino terminus as analyzed by phospholipid affinity column chromatography J Biol Chem 264, 6948–6955 Seemann J, Weber K & Gerke V (1996) Structural requirements for annexin I–S100C complex-formation Biochem J 319, 123–129 Rintala-Dempsey AC, Santamaria-Kisiel L, Liao Y, Lajoie G & Shaw GS (2006) Insights into S100 target specificity examined by a new interaction between S100A11 and annexin A2 Biochemistry 45, 14695– 14705 ´ ´ Rety S, Osterloh D, Arie JP, Tabaries S, Seeman J, Russo-Marie F, Gerke V & Lewit-Bentley A (2000) Structural basis of the Ca2+-dependent association between S100C (S100A11) and its target, the N-terminal part of annexin I Structure 8, 175–184 Filipek A & Wojda U (1996) p30, a novel protein target of mouse calcyclin (S100A6) Biochem J 320, 585–587 Semov A, Moreno MJ, Onichtchenko A, Abulrob A, Ball M, Ekiel I, Pietrzynski G, Stanimirovic D & Alakhov V (2005) Metastasis-associated protein S100A4 induces angiogenesis through interaction with Annexin II and accelerated plasmin formation J Biol Chem 280, 20833–20841 Chang N, Sutherland C, Hesse E, Winkfein R, Wiehler WB, Pho M, Veillette C, Li S, Wilson DP, Kiss E et al (2007) Identification of a novel interaction between the Ca2+-binding protein S100A11 and the Ca2+- and phospholipid-binding protein annexin A6 Am J Physiol Cell Physiol 292, C1417–C1430 Johnstone SA, Hubaishy I & Waisman DM (1993) Regulation of annexin I-dependent aggregation of phospholipid vesicles by protein kinase C Biochem J 294, 801–807 Zhao XQ, Naka M, Muneyuki M & Tanaka T (2000) Ca2+-dependent inhibition of actin-activated myosin ATPase activity by S100C (S100A11), a novel member of the S100 protein family Biochem Biophys Res Commun 267, 77–79 Hirschhorn RR, Aller P, Yuan ZA, Gibson CW & Baserga R (1984) Cell-cycle-specific cDNAs from mammalian cells temperature sensitive for growth Proc Natl Acad Sci USA 81, 6004–6008 Tomas A & Moss SE (2003) Calcium- and cell cycledependent association of annexin 11 with the nuclear envelope J Biol Chem 278, 20210–20216 Kuznicki J, Filipek A, Heimann P, Kaczmarek L & ´ ´ Kaminska B (1989) Tissue specific distribution of calcyclin – 10.5 kDa Ca2+-binding protein FEBS Lett 254, 141–144 4954 68 Kuznicki J, Kordowska J, Puzianowska M & Woznie´ ´ wicz BM (1992) Calcyclin as a marker of human epithelial cells and fibroblasts Exp Cell Res 200, 425–430 69 Filipek A, Puzianowska M, Cies´ lak B & Kuznicki J ´ (1993) Calcyclin – Ca2+-binding protein homologous to glial S-100 beta is present in neurones Neuroreport 4, 383–386 70 Stradal TB & Gimona M (1999) Ca2+-dependent association of S100A6 (Calcyclin) with the plasma membrane and the nuclear envelope J Biol Chem 274, 31593–31596 71 Williams LH, McClive PJ, Van Den Bergen JA & Sinclair AH (2005) Annexin XI co-localises with calcyclin in proliferating cells of the embryonic mouse testis Dev Dyn 234, 432–437 72 Calabretta B, Battini R, Kaczmarek L, de Riel JK & Baserga R (1986) Molecular cloning of the cDNA for a growth factor-inducible gene with strong homology to S-100, a calcium-binding protein J Biol Chem 261, 12628–12632 73 Tonini GP, Fabretti G, Kuznicki J, Massimo L, Scaruffi P, Brisigotti M & Mazzocco K (1995) Gene expression and protein localisation of calcyclin, a calcium-binding protein of the S-100 family in fresh neuroblastomas Eur J Cancer 31A, 499–504 74 Guo XJ, Chambers AF, Parfett CL, Waterhouse P, Murphy LC, Reid RE, Craig AM, Edwards DR & Denhardt DT (1990) Identification of a serum-inducible messenger RNA (5B10) as the mouse homologue of calcyclin: tissue distribution and expression in metastatic, ras-transformed NIH 3T3 cells Cell Growth Differ 1, 333–338 75 Tonini GP, Casalaro A, Cara A & Di Martino D (1991) Inducible expression of calcyclin, a gene with strong homology to S-100 protein, during neuroblastoma cell differentiation and its prevalent expression in Schwann-like cell lines Cancer Res 51, 1733–1737 76 Tokumitsu H, Mizutani A, Minami H, Kobayashi R & Hidaka H (1992) A calcyclin-associated protein is a newly identified member of the Ca2+ ⁄ phospholipidbinding proteins, annexin family J Biol Chem 267, 8919–8924 77 Tokumitsu H, Kobayashi R & Hidaka H (1991) A calcium-binding protein from rabbit lung cytosol identified as the product of growth-regulated gene (2A9) and its binding proteins Arch Biochem Biophys 288, 202– 207 78 Kuznicki J & Filipek A (1987) Purification and ´ properties of a novel Ca2+-binding protein (10.5 kDa) from Ehrlich-ascites-tumour cells Biochem J 247, 663– 667 79 Zimmer DB & Landar A (1995) Analysis of S100A1 expression during skeletal muscle and neuronal cell differentiation J Neurochem 64, 2727–2736 FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS N Miwa et al 80 Zimmer DB, Cornwall EH, Landar A & Song W (1995) The S100 protein family: history, function, and expression Brain Res Bull 37, 417–429 81 Kaetzel MA, Pula G, Campos B, Uhrin P, Horseman N & Dedman JR (1994) Annexin VI isoforms are differentially expressed in mammalian tissues Biochim Biophys Acta 1223, 368–374 82 Arcuri C, Giambanco I, Bianchi R & Donato R (2002) Annexin V, annexin VI, S100A1 and S100B in developing and adult avian skeletal muscles Neuroscience 109, 371–388 83 Garbuglia M, Verzini M & Donato R (1998) Annexin VI binds S100A1 and S100B and blocks the ability of S100A1 and S100B to inhibit desmin and GFAP assemblies into intermediate filaments Cell Calcium 24, 177– 191 84 Garbuglia M, Verzini M, Hofmann A, Huber R & Donato R (2000) S100A1 and S100B interactions with annexins Biochim Biophys Acta 1498, 192–206 Biology of S100–annexin complexes 85 Bianchi R, Verzini M, Garbuglia M, Giambanco I & Donato R (1994) Mechanism of S100 protein-dependent inhibition of glial fibrillary acidic protein (GFAP) polymerization Biochim Biophys Acta 1223, 354–360 86 Garbuglia M, Verzini M, Giambanco I, Spreca A & Donato R (1996) Effects of calcium-binding proteins (S-100a(o), S-100a, S-100b) on desmin assembly in vitro FASEB J 10, 317–324 87 McNeil PL & Steinhardt RA (2003) Plasma membrane disruption: repair, prevention, adaptation Annu Rev Cell Dev Biol 19, 697–731 88 Zeng FY, Gerke V & Gabius HJ (1993) Identification of annexin II, annexin VI and glyceraldehyde-3-phosphate dehydrogenase as calcyclin-binding proteins in bovine heart Int J Biochem 25, 1019–1027 89 Hatakeyama T, Okada M, Shimamoto S, Kubota Y & Kobayashi R (2004) Identification of intracellular target proteins of the calcium-signaling protein S100A12 Eur J Biochem 271, 3765–3775 FEBS Journal 275 (2008) 4945–4955 ª 2008 The Authors Journal compilation ª 2008 FEBS 4955 ... olfactory and respiratory cilia (see below) Biology of S100–annexin complexes S100–annexin complexes S100A10–annexin A2 complex Distribution The mRNA expression of S100A10 and annexin A2 has been shown... S100A10–annexin A2 complex (see above) Other S100–annexin complexes Although the number of reports is limited, other S10 0– annexin complexes have been reported: S100A4–annexin A2 [61], S100A6–annexin... mechanisms of fibrinolysis Br J Haematol 129, 30 7– 321 Biology of S100–annexin complexes 42 Waisman DM (2005) Annexin A2 may not play a role as a plasminogen receptor Br J Haematol 131, 55 2– 556 43