Abstract
The range of applications for large-scale synthetic molecule libraries (1–7) can be expanded if the constituents can be liberated locally from their supporting matrix in a controlled manner so that fractions are available for multiple independent tests, free of interference from other constituents of the library. A method was developed to study the functional responses arlsing from individual constituent beads in a synthetic combinatorial peptide library by introducing the multiuse peptide library (MUPL) concept (8). In the MUPL method ( Fig. 1 ), peptides are liberated from their supports in a dry state so that the problem of signal interference caused by mixing of peptlde molecules, particularly agonists and antagonists, is avoided. In addition, the peptides are released rn a controlled manner so that fractions are available for repetitive screens, thus elimmating the need for iterative library analysis and resynthesis Since the liberated constituents are not constrained by the tethered linkers, the molecules are free to assume their native conformations. These unique features of an MUPL has enabled the use of large-scale synthetic molecule libraries for functional screening (8–10).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Geysen H. M., Meloen R. H., and Barteling S. J (1984) Use of peptlde synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc. Natl. Acad Scl. USA 81, 3998–4002.
Fodor S P A., Read J L., Pirrung M C., Stryer L., Lu A T., and Solas D. (1991) Lrght-directed, spatially addressable parallel chemical synthesis Science 251, 767–773
Furka A., Sebestyen F., Asgedom M., and Dibo G (1991) General method for rapid synthesis of multicomponent peptide mrxtures Int J.Pept Protein Res. 37, 487–493.
Lam K. S., Salmon S E., Hersh E. M., Hruby V J., Kazmlerski W M., and Knapp R. J. (1991) A new type of synthetic peptlde library for identifying ligand-binding actrvrty. Nature 354, 82–84
Houghten R A., Pnulla C., Blondelle S E., Appel J R., Dooley C T., and Cuervo J H. (1991) Generation and use of synthetic peptlde combinatorial librartes for basic research and drug discovery. Nature 354, 84–86.
Gallop M A., Barrett R. W., Dower W. J., Fodor S P. A., and Gordon E M. (1994) Applications of combinatorial technologres to drug discovery 1. Background and peptlde combinatorial libraries J.Med Chem. 37, 1233–1251.
Gordon E M., Barrett R. W., Dower W J., Fodor S. P. A., and Gallop M. A (1994) Applications of combinatorial technologtes to drug discovery 2. Combinatorial organic synthesis, library screening strategies, and future directions. J.Med Chem. 37, 1385–1401.
Jayawlckreme C K., Graminski G. F., Quillan J M., and Lerner M R. (1994) Creation and functional screemng of a multi-use peptide library Proc Natl. Acad Scz USA 91, 1614–1618.
Jayawickreme C K., Quillan J M., Graminski G F., and Lerner M R (1994) Discovery and structure-function analysis of α-melanocyte-stimulating hormone antagonists. J. Bzol. Chem. 47, 29,846–29,854
Quillan J M., Jayawickreme C K., and Lerner M. R (1995) Cotnbinatorlal diffusion assay used to ldentlfy topically active melanocyte-stlmulating hormone receptor antagomsts Proc. Natl. Acad. Scl USA 92, 2894–2898
Jayawlckreme C. K and Lerner M. R. (1993) Generation and screening of mobile peptide libraries FASEB J. 7, A1237 (abstract).
Matsueda G R and Stewart J.M. (1981) A p-Methylbenzhydrylamme resin for Improved solid-phase synthesis of peptide amides Peptzdes 2, 45–50.
Fields G B., Tlan Z., and Barany G. (1992) Principles and practice of solid phase peptlde synhesis, in Synthetzc Peptldes: A User’s Guide (Grant G A., ed.), Freeman, New York, pp 77–183
Fields G B and Noble R L. (1990) Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids Znt J.Pept Protein Res. 35, 161–214
Fields C. G., Lloyd D H., Macdonald R. L., Otteson K. M., and Noble R. L. (1991) HBTU activation for automated Fmoc solid-phase peptide synthesis. Pept. Res. 4, 95–101.
Sarin V.K., Kent S.B H., Tam J P., and Merrifield R B (1981) Quantitative monitoring of solid-phase peptide synthesis by the nmhydrin reaction Anal Biochem. 117, 147–157
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Jayawickreme, C.K., Jayawickreme, S.P., Lerner, M.R. (1998). Generation of Multiuse Peptide Libraries for Functional Screenings. In: Cabilly, S. (eds) Combinatorial Peptide Library Protocols. Methods in Molecular Biology™, vol 87. Humana Press. https://doi.org/10.1385/0-89603-392-9:107
Download citation
DOI: https://doi.org/10.1385/0-89603-392-9:107
Publisher Name: Humana Press
Print ISBN: 978-0-89603-392-4
Online ISBN: 978-1-59259-571-6
eBook Packages: Springer Protocols