全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
PLOS ONE  2012 

A Highly Scalable Peptide-Based Assay System for Proteomics

DOI: 10.1371/journal.pone.0037441

Full-Text   Cite this paper   Add to My Lib

Abstract:

We report a scalable and cost-effective technology for generating and screening high-complexity customizable peptide sets. The peptides are made as peptide-cDNA fusions by in vitro transcription/translation from pools of DNA templates generated by microarray-based synthesis. This approach enables large custom sets of peptides to be designed in silico, manufactured cost-effectively in parallel, and assayed efficiently in a multiplexed fashion. The utility of our peptide-cDNA fusion pools was demonstrated in two activity-based assays designed to discover protease and kinase substrates. In the protease assay, cleaved peptide substrates were separated from uncleaved and identified by digital sequencing of their cognate cDNAs. We screened the 3,011 amino acid HCV proteome for susceptibility to cleavage by the HCV NS3/4A protease and identified all 3 known trans cleavage sites with high specificity. In the kinase assay, peptide substrates phosphorylated by tyrosine kinases were captured and identified by sequencing of their cDNAs. We screened a pool of 3,243 peptides against Abl kinase and showed that phosphorylation events detected were specific and consistent with the known substrate preferences of Abl kinase. Our approach is scalable and adaptable to other protein-based assays.

References

[1]  Wetterstrand KA (2011) DNA Sequencing Costs: Data from the NHGRI Large-Scale Genome Sequencing Program. National Human Genome Research Institute website. 24: Available: www.genome.gov/sequencingcosts Accessed 2012 April.
[2]  Finlay WJ, Bloom L, Cunningham O (2011) Phage display: a powerful technology for the generation of high specificity affinity reagents from alternative immune sources. Methods Mol Biol 681: 87–101.
[3]  Whitney M, Crisp JL, Olson ES, Aguilera TA, Gross LA, et al. (2010) Parallel in vivo and in vitro selection using phage display identifies protease-dependent tumor-targeting peptides. J Biol Chem 285: 22532–22541.
[4]  Brown KC (2010) Peptidic tumor targeting agents: the road from phage display peptide selections to clinical applications. Curr Pharm Des 16: 1040–1054.
[5]  Scholle MD, Kriplani U, Pabon A, Sishtla K, Glucksman MJ, et al. (2006) Mapping protease substrates by using a biotinylated phage substrate library. Chembiochem 7: 834–838.
[6]  Dente L, Vetriani C, Zucconi A, Pelicci G, Lanfrancone L, et al. (1997) Modified phage peptide libraries as a tool to study specificity of phosphorylation and recognition of tyrosine containing peptides. J Mol Biol 269: 694–703.
[7]  Matthews DJ, Wells JA (1993) Substrate phage: selection of protease substrates by monovalent phage display. Science 260: 1113–1117.
[8]  Kurz M, Gu K, Al-Gawari A, Lohse PA (2001) cDNA - protein fusions: covalent protein - gene conjugates for the in vitro selection of peptides and proteins. Chembiochem 2: 666–672.
[9]  Roberts RW, Szostak JW (1997) RNA-peptide fusions for the in vitro selection of peptides and proteins. Proc Natl Acad Sci U S A 94: 12297–12302.
[10]  Ueda T, Kanamori T, Ohashi H (2010) Ribosome display with the PURE technology. Methods Mol Biol 607: 219–225.
[11]  Bertschinger J, Grabulovski D, Neri D (2007) Selection of single domain binding proteins by covalent DNA display. Protein Eng Des Sel 20: 57–68.
[12]  Sergeeva A, Kolonin MG, Molldrem JJ, Pasqualini R, Arap W (2006) Display technologies: application for the discovery of drug and gene delivery agents. Adv Drug Deliv Rev 58: 1622–1654.
[13]  de Figueiredo P, Roberts RL, Nester EW (2004) DARTs: A DNA-based in vitro polypeptide display technology. Proteomics 4: 3128–3140.
[14]  Odegrip R, Coomber D, Eldridge B, Hederer R, Kuhlman PA, et al. (2004) CIS display: In vitro selection of peptides from libraries of protein-DNA complexes. Proc Natl Acad Sci U S A 101: 2806–2810.
[15]  Yonezawa M, Doi N, Kawahashi Y, Higashinakagawa T, Yanagawa H (2003) DNA display for in vitro selection of diverse peptide libraries. Nucleic Acids Res 31: e118.
[16]  Hanes J, Plückthun A (1997) In vitro selection and evolution of functional proteins by using ribosome display. Proc Natl Acad Sci U S A 94: 4937–4942.
[17]  Ullman CG, Frigotto L, Cooley RN (2011) In vitro methods for peptide display and their applications. Brief Funct Genomics 10: 125–134.
[18]  Yamagishi Y, Shoji I, Miyagawa S, Kawakami T, Katoh T, et al. (2011) Natural Product-Like Macrocyclic N-Methyl-Peptide Inhibitors against a Ubiquitin Ligase Uncovered from a Ribosome-Expressed De Novo Library. Chem Biol 18: 1562–1570.
[19]  Xu L, Aha P, Gu K, Kuimelis RG, Kurz M, et al. (2002) Directed evolution of high-affinity antibody mimics using mRNA display. Chem Biol 9: 933–942.
[20]  Kong Y (2009) Calculating complexity of large randomized libraries. J Theor Biol 259: 641–645.
[21]  Szardenings M (2003) Phage display of random peptide libraries: applications, limits, and potential. J Recept Signal Transduct Res 23: 307–349.
[22]  Keefe AD, Szostak JW (2001) Functional proteins from a random-sequence library. Nature 410: 715–718.
[23]  Shivange AV, Marienhagen J, Mundhada H, Schenk A, Schwaneberg U (2009) Advances in generating functional diversity for directed protein evolution. Curr Opin Chem Biol 13: 19–25.
[24]  Takahashi TT, Roberts RW (2009) In vitro selection of protein and peptide libraries using mRNA display. Methods in molecular biology (Clifton, NJ) 535: 293.
[25]  Overall CM, Blobel CP (2007) In search of partners: linking extracellular proteases to substrates. Nat Rev Mol Cell Biol 8: 245–257.
[26]  Rawlings ND, Morton FR, Barrett AJ (2006) MEROPS: the peptidase database. Nucleic Acids Res 34: D270–D272.
[27]  Lopez-Otin C, Overall CM (2002) Protease degradomics: a new challenge for proteomics. Nat Rev Mol Cell Biol 3: 509–519.
[28]  Diks SH, Parikh K, van der Sijde M, Joore J, Ritsema T, et al. (2007) Evidence for a minimal eukaryotic phosphoproteome? PLoS One 2: e777.
[29]  Manning G, Whyte DB, Martinez R, Hunter T, Sudarsanam S (2002) The protein kinase complement of the human genome. Science 298: 1912–1934.
[30]  Chong PK, Lee H, Kong JW, Loh MC, Wong CH, et al. (2008) Phosphoproteomics, oncogenic signaling and cancer research. Proteomics 8: 4370–4382.
[31]  Johnson SA, Hunter T (2005) Kinomics: methods for deciphering the kinome. Nat Methods 2: 17–25.
[32]  Manning G, Plowman GD, Hunter T, Sudarsanam S (2002) Evolution of protein kinase signaling from yeast to man. Trends Biochem Sci 27: 514–520.
[33]  Vasiljeva O, Reinheckel T, Peters C, Turk D, Turk V, et al. (2007) Emerging roles of cysteine cathepsins in disease and their potential as drug targets. Curr Pharm Des 13: 385–401.
[34]  Winssinger N, Damoiseaux R, Tully DC, Geierstanger BH, Burdick K, et al. (2004) PNA-encoded protease substrate microarrays. Chem Biol 11: 1351–1360.
[35]  Collins MO, Yu L, Choudhary JS (2007) Analysis of protein phosphorylation on a proteome-scale. Proteomics 7: 2751–2768.
[36]  Villén J, Beausoleil SA, Gerber SA, Gygi SP (2007) Large-scale phosphorylation analysis of mouse liver. Proc Natl Acad Sci U S A 104: 1488–1493.
[37]  Parikh K, Peppelenbosch MP (2010) Kinome profiling of clinical cancer specimens. Cancer Res 70: 2575–2578.
[38]  Piersma SR, Labots M, Verheul HM, Jiménez CR (2010) Strategies for kinome profiling in cancer and potential clinical applications: chemical proteomics and array-based methods. Anal Bioanal Chem 397: 3163–3171.
[39]  Paradela A, Albar JP (2008) Advances in the analysis of protein phosphorylation. J Proteome Res 7: 1809–1818.
[40]  Rush J, Moritz A, Lee KA, Guo A, Goss VL, et al. (2005) Immunoaffinity profiling of tyrosine phosphorylation in cancer cells. Nat Biotechnol 23: 94–101.
[41]  Rothman DM, Shults MD, Imperiali B (2005) Chemical approaches for investigating phosphorylation in signal transduction networks. Trends Cell Biol 15: 502–510.
[42]  Schutkowski M, Reineke U, Reimer U (2005) Peptide arrays for kinase profiling. Chembiochem 6: 513–521.
[43]  Bonetta (2005) Probing the kinome. Nature Methods 2: 225–232.
[44]  Choudhary C, Mann M (2010) Decoding signalling networks by mass spectrometry-based proteomics. Nat Rev Mol Cell Biol 11: 427–439.
[45]  Yen M, Yin J (2007) High-throughput profiling of posttranslational modification enzymes by phage display. Biotechniques 43: 31, 33, 35 passim.
[46]  Larman HB, Zhao Z, Laserson U, Li MZ, Ciccia A, et al. (2011) Autoantigen discovery with a synthetic human peptidome. Nat Biotechnol 29: 535–541.
[47]  Mahler M, Fritzler MJ (2010) Epitope specificity and significance in systemic autoimmune diseases. Ann N Y Acad Sci 1183: 267–287.
[48]  Pogson M, Georgiou G, Iverson BL (2009) Engineering next generation proteases. Curr Opin Biotechnol 20: 390–397.
[49]  Jung JP, Gasiorowski JZ, Collier JH (2010) Fibrillar peptide gels in biotechnology and biomedicine. Biopolymers 94: 49–59.
[50]  Boschetti E, Righetti PG (2009) The art of observing rare protein species in proteomes with peptide ligand libraries. Proteomics 9: 1492–1510.
[51]  Estephan E, Larroque C, Bec N, Martineau P, Cuisinier FJ, et al. (2009) Selection and mass spectrometry characterization of peptides targeting semiconductor surfaces. Biotechnol Bioeng 104: 1121–1131.
[52]  Whaley SR, English DS, Hu EL, Barbara PF, Belcher AM (2000) Selection of peptides with semiconductor binding specificity for directed nanocrystal assembly. Nature 405: 665–668.
[53]  Lee S, Xie J, Chen X (2010) Peptide-based probes for targeted molecular imaging. Biochemistry 49: 1364–1376.
[54]  Salomon AR, Ficarro SB, Brill LM, Brinker A, Phung QT, et al. (2003) Profiling of tyrosine phosphorylation pathways in human cells using mass spectrometry. Proc Natl Acad Sci U S A 100: 443–448.
[55]  Ryan KJ, Ray CG (2004) Sherris Medical Microbiology. McGraw Hill.
[56]  Kwong AD, Kim JL, Rao G, Lipovsek D, Raybuck SA (1999) Hepatitis C virus NS3/4A protease. Antiviral Res 41: 67–84.
[57]  Kwong AD, Kauffman RS, Hurter P, Mueller P (2011) Discovery and development of telaprevir: an NS3-4A protease inhibitor for treating genotype 1 chronic hepatitis C virus. Nat Biotechnol 29: 993–1003.
[58]  Steuber H, Hilgenfeld R (2010) Recent advances in targeting viral proteases for the discovery of novel antivirals. Curr Top Med Chem 10: 323–345.
[59]  Chen SH, Tan SL (2005) Discovery of small-molecule inhibitors of HCV NS3-4A protease as potential therapeutic agents against HCV infection. Curr Med Chem 12: 2317–2342.
[60]  Lahm A, Yagnik A, Tramontano A, Koch U (2002) Hepatitis C virus proteins as targets for drug development: the role of bioinformatics and modelling. Curr Drug Targets 3: 281–296.
[61]  Hahm B, Han DS, Back SH, Song OK, Cho MJ, et al. (1995) NS3-4A of hepatitis C virus is a chymotrypsin-like protease. J Virol 69: 2534–2539.
[62]  Blight KJ, Kolykhalov AA, Reed KE, Agapov EV, Rice CM (1998) Molecular virology of hepatitis C virus: an update with respect to potential antiviral targets. Antivir Ther 3: 71–81.
[63]  Steinkühler C, Urbani A, Tomei L, Biasiol G, Sardana M, et al. (1996) Activity of purified hepatitis C virus protease NS3 on peptide substrates. J Virol 70: 6694–6700.
[64]  Bartenschlager R, Ahlborn-Laake L, Yasargil K, Mous J, Jacobsen H (1995) Substrate determinants for cleavage in cis and in trans by the hepatitis C virus NS3 proteinase. J Virol 69: 198–205.
[65]  Shiryaev SA, Thomsen ER, Cieplak P, Chudin E, Cheltsov AV, et al. (2012) New details of HCV NS3/4A proteinase functionality revealed by a high-throughput cleavage assay. PLoS ONE 7(4): e35759.
[66]  B?hmer FD, Uecker A (2009) A substrate peptide for the FLT3 receptor tyrosine kinase. Br J Haematol 144: 127–130.
[67]  Jalal S, Arsenault R, Potter AA, Babiuk LA, Griebel PJ, et al. (2009) Genome to kinome: species-specific Peptide arrays for kinome analysis. Sci Signal 2: pl1.
[68]  Amanchy R, Zhong J, Molina H, Chaerkady R, Iwahori A, et al. (2008) Identification of c-Src tyrosine kinase substrates using mass spectrometry and peptide microarrays. J Proteome Res 7: 3900–3910.
[69]  Shults MD, Kozlov IA, Nelson N, Kermani BG, Melnyk PC, et al. (2007) A multiplexed protein kinase assay. Chembiochem 8: 933–942.
[70]  Hornbeck PV, Chabra I, Kornhauser JM, Skrzypek E, Zhang B (2004) PhosphoSite: A bioinformatics resource dedicated to physiological protein phosphorylation. Proteomics 4: 1551–1561.
[71]  Yang CY, Chang CH, Yu YL, Lin TC, Lee SA, et al. (2008) PhosphoPOINT: a comprehensive human kinase interactome and phospho-protein database. Bioinformatics 24: i14–i20.
[72]  Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, et al. (2006) Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell 127: 635–648.
[73]  Dinkel H, Chica C, Via A, Gould CM, Jensen LJ, et al. (2011) Phospho.ELM: a database of phosphorylation sites–update 2011. Nucleic Acids Res 39: D261–D267.
[74]  Rychlewski L, Kschischo M, Dong L, Schutkowski M, Reimer U (2004) Target specificity analysis of the Abl kinase using peptide microarray data. J Mol Biol 336: 307–311.
[75]  Songyang Z, Carraway KL, Eck MJ, Harrison SC, Feldman RA, et al. (1995) Catalytic specificity of protein-tyrosine kinases is critical for selective signalling. Nature 373: 536–539.
[76]  Heibeck TH, Ding SJ, Opresko LK, Zhao R, Schepmoes AA, et al. (2009) An extensive survey of tyrosine phosphorylation revealing new sites in human mammary epithelial cells. J Proteome Res 8: 3852–3861.
[77]  J?rgensen C, Sherman A, Chen GI, Pasculescu A, Poliakov A, et al. (2009) Cell-specific information processing in segregating populations of Eph receptor ephrin-expressing cells. Science 326: 1502–1509.
[78]  Stuible M, Dubé N, Tremblay ML (2008) PTP1B regulates cortactin tyrosine phosphorylation by targeting Tyr446. J Biol Chem 283: 15740–15746.
[79]  Choudhary C, Kumar C, Gnad F, Nielsen ML, Rehman M, et al. (2009) Lysine acetylation targets protein complexes and co-regulates major cellular functions. Science 325: 834–840.
[80]  Uy R, Wold F (1977) Posttranslational covalent modification of proteins. Science 198: 890–896.
[81]  Goto Y, Katoh T, Suga H (2011) Flexizymes for genetic code reprogramming. Nat Protoc 6: 779–790.
[82]  Ohta A, Yamagishi Y, Suga H (2008) Synthesis of biopolymers using genetic code reprogramming. Curr Opin Chem Biol 12: 159–167.
[83]  Frankel A, Millward SW, Roberts RW (2003) Encodamers: unnatural peptide oligomers encoded in RNA. Chem Biol 10: 1043–1050.
[84]  Shiryaev SA, Kozlov IA, Ratnikov BI, Smith JW, Lebl M, et al. (2007) Cleavage preference distinguishes the two-component NS2B-NS3 serine proteinases of Dengue and West Nile viruses. Biochem J 401: 743–752.
[85]  Hattori S, Iida N, Kosako H (2008) Identification of protein kinase substrates by proteomic approaches. Expert Rev Proteomics 5: 497–505.
[86]  Kozlov IA, Melnyk PC, Zhao C, Hachmann JP, Shevchenko V, et al. (2006) A method for rapid protease substrate evaluation and optimization. Comb Chem High Throughput Screen 9: 481–487.
[87]  Peck SC (2006) Analysis of protein phosphorylation: methods and strategies for studying kinases and substrates. Plant J 45: 512–522.
[88]  George J, Teear ML, Norey CG, Burns DD (2003) Evaluation of an imaging platform during the development of a FRET protease assay. J Biomol Screen 8: 72–80.
[89]  Manning BD, Cantley LC (2002) Hitting the target: emerging technologies in the search for kinase substrates. Sci STKE 2002: PE49. PE49 p.
[90]  Stadler V, Felgenhauer T, Beyer M, Fernandez S, Leibe K, et al. (2008) Combinatorial synthesis of peptide arrays with a laser printer. Angew Chem Int Ed Engl 47: 7132–7135.
[91]  Beyer M, Nesterov A, Block I, K?nig K, Felgenhauer T, et al. (2007) Combinatorial synthesis of peptide arrays onto a microchip. Science 318: 1888.
[92]  Hilpert K, Winkler DF, Hancock RE (2007) Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion. Nat Protoc 2: 1333–1349.
[93]  Breitling F, Nesterov A, Stadler V, Felgenhauer T, Bischoff FR (2009) High-density peptide arrays. Mol Biosyst 5: 224–234.
[94]  LeProust EM, Peck BJ, Spirin K, McCuen HB, Moore B, et al. (2010) Synthesis of high-quality libraries of long (150mer) oligonucleotides by a novel depurination controlled process. Nucleic Acids Res 38: 2522–2540.
[95]  Miyamoto-Sato E, Takashima H, Fuse S, Sue K, Ishizaka M, et al. (2003) Highly stable and efficient mRNA templates for mRNA-protein fusions and C-terminally labeled proteins. Nucleic Acids Res 31: e78.
[96]  Pieles U, Englisch U (1989) Psoralen covalently linked to oligodeoxyribonucleotides: synthesis, sequence specific recognition of DNA and photo-cross-linking to pyrimidine residues of DNA. Nucleic Acids Res 17: 285–299.
[97]  Tolbert TJ, Wong CH (2002) New methods for proteomic research: preparation of proteins with N-terminal cysteines for labeling and conjugation. Angew Chem Int Ed Engl 41: 2171–2174.
[98]  Blanco-Canosa JB, Dawson PE (2008) An efficient Fmoc-SPPS approach for the generation of thioester peptide precursors for use in native chemical ligation. Angew Chem Int Ed Engl 47: 6851–6855.
[99]  Bentley DR, Balasubramanian S, Swerdlow HP, Smith GP, Milton J, et al. (2008) Accurate whole human genome sequencing using reversible terminator chemistry. Nature 456: 53–59.
[100]  He M, Stoevesandt O, Palmer EA, Khan F, Ericsson O, et al. (2008) Printing protein arrays from DNA arrays. Nat Methods 5: 175–177.
[101]  Tao SC, Zhu H (2006) Protein chip fabrication by capture of nascent polypeptides. Nat Biotechnol 24: 1253–1254.
[102]  Ramachandran N, Hainsworth E, Bhullar B, Eisenstein S, Rosen B, et al. (2004) Self-assembling protein microarrays. Science 305: 86–90.
[103]  He M, Taussig MJ (2001) Single step generation of protein arrays from DNA by cell-free expression and in situ immobilisation (PISA method). Nucleic Acids Res 29: E73–E73.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133