Quantitative screening of EGF receptor-binding peptides by using a peptide library with multiple fluorescent amino acids as fluorescent tags

Bioorganic and Medicinal Chemistry Letters

Volumn 20, Issue 20, 2010, Pages 5976-5978

  Kitamatsu, Mizuki ^a   Yamamoto, Takahiro ^a   Futami, Midori ^a   Sisido, Masahiko ^a  

^a OKAYAMA UNIVERSITY   (Japan)

Author keywords

EGFR; Fluorescent amino acid; Peptide; Peptide library; Peptide screening

Indexed keywords

AMINO ACID; BINDING PROTEIN; EPIDERMAL GROWTH FACTOR RECEPTOR; PEPTIDE LIBRARY;

ARTICLE; CHEMICAL STRUCTURE; FLUORESCENCE; PEPTIDE SYNTHESIS; PROTEIN BINDING; QUANTITATIVE ANALYSIS; STRUCTURE ACTIVITY RELATION;

AMINO ACID SEQUENCE; AMINO ACIDS; FLUORESCENT DYES; HUMANS; MOLECULAR STRUCTURE; PEPTIDE LIBRARY; PEPTIDES; PROTEIN BINDING; RECEPTOR, EPIDERMAL GROWTH FACTOR;

EID: 77956934165     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.08.078     Document Type: Article

References (23)

1. J.P. Scott, and G.P. Smith Science 249 1990 386
2. K.S. Lam, S.E. Salmon, E.M. Hersh, V.J. Hruby, W.M. Kazmierski, and R.J. Knapp Nature 354 1991 82
3. K.S. Lam, M. Lebl, and V. Krchnak Chem. Rev. 97 1997 411
4. M.M. Marani, M.C. Martínez Ceron, S.L. Giudicessi, E. de Oliveira, S. Côté, R. Erra-Balsells, F. Albericio, O. Cascone, and S.A. Camperi J. Comb. Chem. 11 2009 146
5. M. Kitamatsu, M. Futami, and M. Sisido Chem. Commun. 46 2010 761
6. H. Ogiso, R. Ishitani, O. Nureki, S. Fukai, M. Yamanaka, J.-H. Kim, K. Saito, A. Sakamoto, M. Inoue, M. Shirouzu, and S. Yokoyama Cell 110 2002 775
7. J.G. Paez, P.A. Janne, J.C. Lee, S. Tracy, H. Greulich, S. Gabriel, P. Herman, F.J. Kaye, N. Lindeman, T.J. Boggon, K. Naoki, H. Sasaki, Y. Fujii, M.J. Eck, W.R. Sellers, B.E. Johnson, and M. Meyerson Science 304 2004 1497
8. Z. Li, R. Zhao, X. Wu, Y. Sun, M. Yao, J. Li, Y. Xu, and J. Gu FASEB J. 19 2005 1978
9. C. Buerger, K. Nagel-Wolfrum, C. Kunz, I. Wittig, K. Butz, F. Hoppe-Seyler, and B. Groner J. Biol. Chem. 278 2003 37610
10. C.P. Hart, J.E. Martin, M.A. Reed, A.A. Keval, M.J. Pustelnik, J.P. Northrop, D.V. Patel, and J.R. Grove Cell Signalling 11 1999 453
11. X. Tang, P. Dallaire, D.W. Hoyt, B.D. Sykes, M. O'Connor-McCourt, and B.A. Malcolm J. Biochem. 122 1997 686
12. M. Kitamatsu, T. Kubo, R. Matsuzaki, T. Endoh, T. Ohtsuki, and M. Sisido Bioorg. Med. Chem. Lett. 19 2009 3410
13. M. Kitamatsu, T. Kashiwagi, R. Matsuzaki, and M. Sisido Chem. Lett. 35 2006 301
14. M. Kitamatsu, M. Shigeyasu, M. Saitoh, and M. Sisido Biopolymers (Peptide Sci.) 84 2006 267
15. M. Kitamatsu, M. Shigeyasu, T. Okada, and M. Sisido Chem. Commun. 2004 1208
16. Three fluorescent amino acids (Hmc, Dec and Coc) were newly synthesized (see Supplementary data).
17. 2 in the center of the segment of library moiety because we assumed that molecular fluctuation at their positions is less (the amino acids strongly interact with the target).
18. Fifteen natural amino acids were chosen to assign fifteen fluorescent amino acids. Of twenty natural amino acids, Cys was removed to avoid intracyclization, dimerization or polymerization of peptides. Glu, Gly, Leu and Gln were removed to have amino acids of a similar property (Asp, Ala, Ile and Asn, respectively).
19. We selected d-configurated amino acids as the units composing fluorescent peptides because we assumed that resistance to proteolytic degradation of d-configurated peptides is superior to that of l-configurated peptides. We will utilize EGFR-binding peptides found by this method as medical tools in the future.
20. A 100-μL mixture of fluorescent peptides (concentration of each peptide being 5-15 nM) was measured by high-sensitive 2D-FL spectroscopy with a CCD camera (Photon design, Japan).
21. sErbB1 was obtained from sErbB1/293H cells provided by Professor Masaharu Seno, Okayama University.
22. Negative values may be experimental errors ascribed to 2D-FL spectrometer. The errors may be caused by deviations between component spectra and each spectrum of fluorescent peptides obtained by decomposing the objective spectrum.
23. Abbreviations used are as follows: Fmoc, 9-fluorenylmethoxycarbonyl; Hmc, N - (7-hydroxy-4-methyl-coumarin-3-acetyl)-l-lysine; Cm3, N-δ-(coumarin 343-3-carbonyl)-l-ornithine; Moc, N - (7-methoxycoumarin-4-carbonyl)-l-lysine; Coc, N-δ-(6-oxo-6H-[1,3]dioxolo[4,5-g]chromen-8-acetyl)-l-ornithine; Hoc, N - (7-hydroxycoumarine-4-carbonyl)-l-lysine; Cmr, N - (7-methoxycoumarin-4- acetyl)-l-lysine; Mac, N - (7-dimethylaminocoumarin-4-acetyl)-l-lysine; Acm, N-δ-(9-oxoacridin-10(9H)-acetyl)-l-ornithine; @52, N-δ-{(Z)-N-(6- (ethylamino)-2,7,dimethyl-3H-xanthen-3-ylidene-9-ethylcarbonyl)}-l-ornithine; Acd, β-{2-[9(10H)-acridonyl]}-l-alanine; Dec, N - (7-diethylaminocoumarin- 3-carbonyl)-l-lysine; S39, N - (4-(2-(chroman-6-yl)oxazol-5-yl)-1- benzylpyridinium-3-carbonyl)-l-lysine; A43, N - {(8,8-dimethyl-2-oxo-4- (trifluoromethyl)-8,9-dihydro-2H-pyrano[3,2-g]quinolin-6-yl)methanesulfonic acid-9-pentylcarbonyl}-l-lysine; Fam, (fluorescein-5(6)-carbonyl)-l-lysine; Tmr, (tetramethylrhodamine-5(6)-carbonyl)-l-lysine.