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Fig. 1. Photoaffinity labeling. Fig. 2A. CNBr digestion.Fig. 2B. Enzymatic digestion.to Glu C enzymatic digestion that should highlight larger migrated fragments (seeFigure 2B). Again this digestion showed an elevation of 5 to 6 kDa on SDS-PAGEelectrophoresis. We can also exclude intramolecular cross-linking due to the fact that onlythe intracellular fragment of TMD 4 could account for the CNBr digestion’s gain in mass,yet according to molecular models, this region is over 20 Å away from the Bpa and thus toofar to form a covalent bond even at extreme temperatures [5].We have shown that the expression of hAT 1 containing the non-canonical amino acidBpa can be accomplished with reasonable yields. These receptors can easily be identifiedusing photoaffinity labeling. We are also able to identify an auxiliary hAT 1 binding protein,which has at least one CNBr digestion fragment of 1 to 2 kDa and a Glu C digestionfragment of 5 to 6 kDa. Subsequent immunoblotting experiments could be used topositively identify this hAT 1 binding protein. Furthermore, this method can easily beextrapolated to use other non-canonical amino acids that have been developed by the P.G.Schultz group by using the same tRNA and mutated receptor sequences.AcknowledgmentsWe thank Marie-Reine Lefebvre for her expert knowledge and expertise. Funding was supported bythe CIHR.References1. Liu, W., et al. Nature Methods 4(3), 239-244 (2007).2. Perodin, J., et al. Biochemistry 41(48), 14348-14356 (2002).3. Rihakova, L., et al. J. Recept Signal Transduct. Res. 22(1-4), 297-313 (2002).4. Fillion, D., et al. J. Med. Chem. 55(5), 2073-2075 (2010).5. Arsenault, J., et al. Biochem. Pharmacol. 80(7), 990-999 (2010).547
Proceedings of the 31 st European Peptide SymposiumMichal Lebl, Morten Meldal, Knud J. Jensen, Thomas Hoeg-Jensen (Editors)European Peptide Society, 2010The Synthesis of Some Peptides Intended to Be Inhibitors of theRNA-Polymerase of Influenza A VirusOleg V. Matusevich 1 and Oleg I. Kiselev 21 Saint-Petersburg State University, Chemical Faculty, St. Petersburg, 198504, Russia;2 Russian Academy of Medical Sciences, Research Institute of Influenza,St. Petersburg, 197376, RussiaIntroductionThe limited choice of anti-viral drugs is the reason of low efficiency of acute and persistentinfection treatment [1]. The major drawback of modern anti-viral drugs is lowvirus-specific targeting [2]. Considerable possibilities in drug design are connected withdevelopment of peptides homologous to functional domains of virus-specific proteins [3].The work presented here is concerned with the synthesis of some peptides appearingto be the potential inhibitors of the RNA-polymerase of influenza A virus. The effect ofthese peptides is based on the competitive inhibition of virus RNA-polymerase complexorganization, said RNA-polymerase consisting of three subunits, namely PB1, PB2, and PA[4].Results and DiscussionThe choice of peptides to synthesize was premised on the data both on tertiary structure ofthe complex and from biological experiments on strains of Hong Kong isolate andCalifornian isolate. The former is the ancestor of the world outbreak of bird flu while thelatter is known as swine flu and caused boom in 2009.Sixteen fragments from the PB1 subunit of said viruses RNA-polymerase weresynthesized, namely 1-5, 1-20, 1-25, 6-13, 6-25, 14-25, 26-30, 271-290, 381-386, 381-390,391-400, 381-400, 395-400, 411-420, 525-530, and 525-535 fragments. The amino acidsequences of these fragments are represented in the table below.N Designation Amino Acid Sequence1 PB1 (1-5) H-Met-Asp-Val-Asn-Pro-OH2 PB1 (1-20) H-Met-Asp-Val-Asn-Pro-||-Thr-Leu-Leu-Phe-Leu-Lys-Val-Pro-Ala-Gln-Asn-Ala-Ile-Ser-Thr-OH3 PB1 (1-25) H-Met-Asp-Val-Asn-Pro-||-Thr-Leu-Leu-Phe-Leu-Lys-Val-Pro-||-Ala-Gln-Asn-Ala-Ile-Ser-Thr-Thr-Phe-Pro-Tyr-Thr-OH4 PB1 (6-13) H-Thr-Leu-Leu-Phe-Leu-Lys-Val-Pro-OH5 PB1 (6-25) H-Thr-Leu-Leu-Phe-Leu-Lys-Val-Pro-||-Ala-Gln-Asn-Ala-Ile-Ser-Thr-Thr-Phe-Pro-Tyr-Thr-OH6 PB1(14-25) H-Ala-Gln-Asn-Ala-Ile-Ser-Thr-Thr-Phe-Pro-Tyr-Thr-OH7 PB1 (26-30) H-Gly-Asp-Pro-Pro-Tyr-OH8 PB1(271-290) H-Leu-Pro-Val-Gly-Gly-Asn-Glu-Lys-Lys-Ala-Lys-Leu-Ala-Asn-Val-Val-Arg-Lys-Met-Met-OH9 PB1 (381-386) H-Phe-Asn-Glu-Ser-Thr-Arg-OH10 PB1(381-390) H-Phe-Asn-Glu-Ser-Thr-Arg-Lys-Lys-Ile-Glu-OH11 PB1 (381-400) H-Phe-Asn-Glu-Ser-Thr-Arg-||-Lys-Lys-Ile-Glu-Lys-Ile-Arg-Pro-||-Leu-Leu-Val-Glu-Gly-Thr-OH12 PB1(391-400) H-Lys-Ile-Arg-Pro-Leu-Leu-Val-Glu-Gly-Thr-OH13 PB1 (395-400) H-Leu-Leu-Val-Glu-Gly-Thr-OH14 PB1 (411-420) H-Met-Phe-Asn-Met-Leu-Ser-Thr-Val-Leu-Gly-OH15 PB1 (525-530) H-Ile-Gly-Val-Thr-Val-Ile-OH16 PB1 (525-535) H-Ile-Gly-Val-Thr-Val-Ile-Lys-Asn-Asn-Met-Ile-OH548
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Peptides 2010Tales of PeptidesProce
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Peptides 2010Tales of PeptidesProce
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IntroductionWe had the distinct hon
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Scientific programIn planning the 3
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31 st EUROPEAN PEPTIDE SYMPOSIUMSep
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published by John Wiley & Sons as a
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The Josef Rudinger AwardThis award
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Young Investigators' SymposiumThe y
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xxiv
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Design of Peptidyl-Inhibitors for G
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Synthetic Antifreeze Glycopeptide A
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Synthesis and Oxidative Folding of
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Convergent Syntheses of Huprp106-12
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Bactericidal Activity of Small Beta
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Bradykinin Analogues Acylated on Th
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Antimicrobial Activity of Small 3-(
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Interaction of Curcumin with A-Synu
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Fluorescent and Luminescent Fusion
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Cellular Expression of the Human An
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2D IR Spectroscopy of Oligopeptides
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large, non-redundant subset of prot
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The aberrantly glucosylated peptide
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Table 1. Proline cis/trans ratios o
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Table 1. Yields, UV-vis absorption
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Table 1. Purity of the targeted tri
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processes are blocked. Interestingl
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(tb4 n ,1 m )MTII(tb1 n ,4 m )MTIIF
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Fig. 2. a) Part of the 400 MHz HR-M
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Once printed, the amino acid partic
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A) PSA, few seconds73B) PSA, 45 min
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short β strands. In contrast, nume
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neg. control Cs9-Rhd MM-218Fig. 2.
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Table 1. The general structure of t
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% ID in the liver 1 h p.i.100908070
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Peptidyl phosphoranes were first re
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obtained from the same sardines and
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MccJ25 variants were produced by si
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Fig. 2. Proposed signaling mechanis
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Table 1. mCD4-HS12 antiviral activi
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Proceedings of the 31 st European P
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Subject index(S)-2-(1-adamantyl)gly
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chip 18chiral triazine condensing r
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heat stress 348helical conformation
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O-acyl isopeptide method 112obesity
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SH2-ligands 210short collagen-relat
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Author indexAboye, Teshome Leta 142
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Chi, Hongfang 480Chiche, Laurent 6C
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Geronikaki, Athina 444Gessmann, Ren
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Kostova, Kalina 528Kotzia, Georgia
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Nagata, Koji 162Nagayev, Igor Yu. 5
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Salvarese, Nicola 518Sammet, Benedi
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Vauquelin, Georges 138Veiga, Ana Sa
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