... the COUNTESS MR HARPIN, receiver of taxes, also in love with the COUNTESS MR BOBINET, tutor to the COUNT JEANNOT, servant to MR THIBAUDIER CRIQUET, servant to the COUNTESS THE COUNTESSOF ESCARBAGNAS ... the closet, I mean COUN Call in that rascal of a page AND I say, Criquet! COUN Cease that "Criquet" of yours, stupid, and call out "Page." AND Page then, and not Criquet, come and speak to missis ... must be deaf Criq Page! page! SCENE VI. THE COUNTESS, JULIA, ANDRÉE, CRIQUET CRI What is it you want? COUN Where were you, you rascal? CRI In the street, Ma'am COUN Why in the street? CRI You told...
... C rproErv -C CT 29 kDa 24 kDa rmErv -C CT 20 kDa M 45 30 25 20 15 10 CC B rproErv -C+ CT rproErv -C CT rmErv -C+ CT rmErv -C CT 2 Fig (A) Time course of activation to the mature form (I) rproErv -C+ CT ... determined kcat (s)1)a rproErv -C+ CT rproErv-CDCT Latex Erv -C [17] Km (lM)a kcat ⁄ Km (s)1ÆmM)1) Speci c activity at 37 °Cb (UÆmg)1) Speci c activity at 65 °Cb (UÆmg)1) IC50 against E-64 (nM )c 0.0170 ... mature catalytic domain is ˚ 1037 A2, which is 56% of the total surface area of the CT-extension The Leu side chain at the position 23 of the CT-extension occupies specificity pocket S2 of the catalytic...
... activation of a family of cytosolic cysteine proteases, caspases, which are required for many of the morphological changes that occur during apoptosis The mitochondrial release of cytochrome c ... final concentration of 1% Extracts were incubated for 20 at C and centrifuged at 18 000 g in a microcentrifuge at C For immunoprecipitation assays, extracts of transfected cells were immunprecipitated ... transfected in HEK-293T cells led to increased caspase activity When GFP–Itch was cotransfected with tBid, caspase activity was dramatically reduced (Fig 2B, right panel, Itch) In contrast, reducing...
... Countessof Albany, by Violet Paget (AKA Vernon Lee) CHAPTER XI CHAPTER XII CHAPTER XIII CHAPTER XIV CHAPTER XV CHAPTER XVI CHAPTER XVII CHAPTER XVIII CHAPTER XIX CHAPTER XX Chapters Countess ... heir of the Countessof Albany's heir Fabre Now in the possession of Mrs Horace Walpole, of Heckfield Place, Winchfield, Hants LOUISE, COUNTESSOF ALBANY From a pastel once in the possession of ... weary of the life of melancholy solitude, of debauched squalor, of the moral pig-stye in which he had been rotting so many years, the idea of decency, of dignity, of society, of a wife and children...
... specifically recognize cisplatin-damaged DNA: a clue to anticancer activity of cisplatin FASEB J 12, 791–799 30 Kasparkova J & Brabec V (1995) Recognition of DNA interstrand cross-links of cis-diamminedichloroplatinum(Ii) ... p73 DNA recognition of the protein than for its ‘activated’ forms [33] Recently, it has been reported that accessibility of the p53 CTDBS is critical for (sequence-nonspeci c) cisPtDNA recognition ... 1–68 Academic Press Inc., San Diego 28 Takahara PM, Rosenzweig AC, Frederick CA & Lippard SJ (1995) Crystal structure of double-stranded DNA containing the major adduct of the anticancer drug cisplatin...
... for IC and IC–CPY IC (A) or IC–CPY (B), the final concentration of which was lM, was added to PCbased liposomes (0.5 mgÆmL)1 of PE ⁄ PC, PC, PS ⁄ PC, and PtdIns ⁄ PC) in 20 mM Hepes (pH 7.2) containing ... detectable Proteins Liposomes ka (102 d1–7IC PE ⁄ PC PC PS ⁄ PC PtdIns ⁄ PC PG ⁄ PC PtdIns(3)P ⁄ PC PtdIns(3,4)P2 ⁄ PC PtdIns(3,4,5)P3 ⁄ PC PE ⁄ PC PC PS ⁄ PC PtdIns ⁄ PC PG ⁄ PC PtdIns(3)P ⁄ PC ... cytoplasmic localization of IC in the logarithmic-phase cells Previous work on the inhibitory properties of IC in vitro, in which IC was shown to inactivate and interact with CPY under acidic...
... F1a, 5¢-GGACTCAAGAGAGGAACCTG-3¢; F1b, 5¢-CT GCCTTGCTCCACACCTG-3¢; F 1c, 5¢-CTGCCGTCTCC GCCGCCACT-3¢; FU, 5¢-TCAAGCCCCGCTACATAG TT-3¢; and R3, 5¢-AGGAACCAAACACCAAGTGG-3¢ (Fig 2A) Luciferase promoter ... sequence (GenBank accession number NM006853 and NM14497): R1, 5¢-ATGGTGTCTGTGATGTTGCCG-3¢ and R2, 5¢-TTCTCACACTTCTGGTGCTC-3¢ DNA sequences of the PCR products were determined using an automatic DNA ... (primers 5¢-TCATGCTGGTTGAGACTG GA-3¢ and 5¢-CCAGGTGTGGAGCAAGGCAG-3¢) and a 938 bp fragment of promoter (primers, 5¢-CTCACAG CAGCCAGTAAGTG-3¢ and 5¢-ACTCACAGGCTCTGG GGCTG-3¢) were amplified using...
... ATnI1F (5¢-CATATCACCATGGGTTCCCTTG-3¢) and ATnI292R (5¢-CTTGATTTGGATCCTTTAAGGTA TAGC-3¢), ATnI1F and ATnI128R (5¢-GTTCCGGATC CTATCTTCTGGCTTCC-3¢), ATnI130F (5¢-GCCAGAA CCATGGCGGAGGAAC-3¢) and ATnI292R, ... RTnI1F (5¢-CAAACCTCACCATGGGAGAT GAAG-3¢) and RTnI181R (5¢-CCCCGGAGCCGGATCC CCAGCCCC-3¢) These primers were designed based on the sequence retrieved from the GenBank database under accession number ... biological significance of the difference in TnI–TnC interactions, we compared the ability of TnC to neutralize the inhibitory effects of the C- terminal fragments in the presence and absence of Ca2+...
... 5¢-GGACGATGCCACCAGTGCCCTG GACGCCGAGTGCG-3¢ and 5¢-CGCACTCGGCGTC CAGGGCACTGGTGGCATCGTCC-3¢ and complementary primers 5¢-GGATGAGGCTACCAGTGCCCT GGATGCTGGCAACC-3¢ and 5¢-GGTTGCCAGCATC CAGGGCACTGGTAGCCTCATCC-3¢ ... 5¢-GGATGAGGCTACCAGTAC TCTGGACGCCGAGTGCG-3¢ and 5¢-CGCACTCGGC GTCCAGAGTACTGGTAGCCTCATCC-3¢ All primers were purchased from ARK/Sigma The chimeric TAP construct 1V2 was created by ligation of the 1.6 ... 5¢-GGATGAGGCTACCAGTGC TC TGGACGCCTAG TGCGAGCAGGC-3¢ and 5¢-GCCTGCTCGCACTAGG CGTCCAGAGCACTGGTAGCCTCATCC-3¢ All TAP constructs were transfected into T2 cells by electroporation using a Bio-Rad gene...
... chain factors, eukaryotic class polypeptide chain release factor (eRF1) and eukaryotic class polypeptide chain release factor (eRF3) The major functions of eRF1 include recognition of each of the ... structure and function of the eRF1 C- domain A B C E D Fig The solution structure of the C- domain (A) Stereo view of the ensemble of the final 48 calculated structures Twenty-four structures of ... gene that enhances the termination efficiency by stimulating the activity of eRF1 [1–4] eRF1 contains three structurally separated domains, each of which can be assigned a speci c function The N-terminal...
... 5¢-TTCGA GGCCCGCGCCGCAATTGGGGGCCCAGAA-3¢ and 5¢TTCTGGGCCCCCAATTGCGGCGCGGGCCTCGAA-3¢ for E190A, 5¢-ATTCCGGTTACTTTCGCGGCCCGCGC CCAAATT-3¢ and 5¢-AATTTGGGCGCGGGCCGCGA AAGTAACCGGAAT-3¢ for E190A, 5¢-CAAATTGGGGG ... CCCAGCAGCTGGGAAGTCTGAA-3¢ and 5¢-TTCAGA CTTCCCAGCTGCTGGGCCCCCAATTTG-3¢ for E199A, and 5¢-GAAGCTGGGAAGTCTGCACAGTCTGGAGCC AAG-3¢ and 5¢-CTTGGCTCCAGACTGTGCAGACTTCC CAGCTTC-3¢ for E204A Mutated codons ... primers 5¢-TCTCGGAGATCC GACAGA-3¢ and 5¢-CTTTCGGGCTTTGTTAGCAG-3¢, respectively CD spectra were acquired on a J-810 spectropolarimeter (Jasco, Tokyo, Japan) with an attached Peltier temperaturecontrolled...
... dimeric non-mutated CAC, is important in the virus cycle, as confirmed by structural studies of CAC in the presence of various molecules and agents [28–31] Results Relaxation measurements of CACW40A ... feature of the whole dimeric CAC domain Dynamics of monomeric CAC Model-free analysis versus spectral density mapping Our results indicate that the relaxation data of CACW40A could not be satisfactorily ... NMR concentration by using Centriprep Amicon devices (Millipore), with a molecular mass cut-off of 3500 Da Protein structure calculations The determination of the solvent-accessible surface area...
... La Jolla, CA) (18 cycles: 94 C for min, 55 C for min, 72 C for min, followed by a single incubation at 72 C for 10 min) using primer P1, 5¢-CGCCA AGATTCCTCTGGGAACTCCAGCGACAGT-3¢ (nucleotides ... is catalytically inactive and specifically present in the nucleus of transfected COS-7 cells In contrast, all three forms carrying a complete catalytical domain are catalytically active and localized ... mGRK6-A cDNA by PCR (30 cycles: 94 C for min, 70 C or 62 for min, 72 C for min, followed by a single incubation at 72 C for 10 min) using primer P1, 5¢-AGCCCATGGAGCTCGAGAACA TCGTA-3¢ (nucleotides...
... 5-ATggAgCAgAAACTC ATCTCTgAA-3¢ and 5¢-TTCTgCAgCTAACCAATTTTg CTgCCTTTgTT-3¢ for I7, 5-CgggATCCATgCAgCCA gAATCTggggCC-3¢ and 5¢-CCgCTCgAgTCAAgCCAg TgACCgCCTCCC-3¢ for OR17-40 Each PCR consisted of 40 cycles: ... engineering chimeric constructs with only part of the coding sequences of olfactory receptors [3,24] Only the c- myc tag was added at the 5¢-terminus of I7 sequence Olfactory receptor speci c and dose-dependent ... ODORA cells, only 1& of the cells yielded a discrete punctuate labeling at the level of the plasmic membrane (Fig 2C) Intracellular calcium assay Characteristics of the calcium response to odorant...
... MALDI-TOF-TOF mass spectrometry A selection of mass spectra illustrates how the comparison of chymotryptic peptides from different cross-linked complexes and protein controls allows the detection of ... binding of Ssa1p in the vicinity of this stretch interferes with Ure2p assembly into fibrils either because of a change in the conformation of this stretch or the crowding of a surface interface involved ... MS Covalent cross-linking approaches allow: (a) the identification of surface areas involved in protein-protein interactions within protein complexes; (b) the characterization of the distance constraints...
... amplification of a 70 bp PCR product with [5¢-32P]-labeled S70ds ⁄ UP (5¢-AATTGATCCCGCCCGCCTC-3¢) and S70ds ⁄ DN (5¢-GCCCCG CAGCGGCGCCATGTT-3¢) primers For PCR, we added 2.5 · 10)3 pmol of primer ... (5¢-AATTGATCCCGCCCG CCTCGTTTTCATCGATGAGACCTGGACGAAGACGA ACATGGCGCCGCTGCGGGGC-3¢) using 50 lCi of [c- 32P]ATP (3000 CiÆmmol)1; GE Healthcare) and 100 units of T4 polynucleotide kinase (New England Biolabs Inc., ... of primers were designed to amplify the 5¢ and 3¢ sections of the IEP, respectively: a 5¢ end primer mut UP (5¢-GTCAGCGGTGCTGGAAG TATG-3¢) and a 3¢ end primer A400V ⁄ DN (5¢-ATTTT CCCGCACCAGCTTTCGCAAGA-3¢)...
... nuclear translocation of the intracellular C- terminal domain (ICD) of the APP family of proteins Biochemistry 42, 6664–6673 Cao X & Sudhof TC (2001) A transcriptionally [correction of transcriptively] ... antiserum C8 , which recognizes the C- terminus of APP, and the polyclonal antiserum D2-II, which recognizes the ectodomain of APLP2 (Calbiochem, EMD Chemicals Inc., Gibbstown, NJ, USA), have been described ... levels of soluble APP remaining constant irrespective of the presence or absence of BACE1 The levels of APPsa increased to account for the loss of APPsb (soluble C- terminally truncated b-cleaved...