The use of synthetic peptide combinatorial libraries for the identification of bioactive peptides

Torrey Pines Institute for Molecular Studies, San Diego, CA 92121.
BioTechniques (Impact Factor: 2.95). 10/1992; 13(3):412-21.
Source: PubMed


The systematic preparation of synthetic peptide combinatorial libraries (SPCLs), each composed of tens of millions of peptides that can be screened in existing diagnostically or pharmacologically relevant in vitro assay systems, is reviewed. The identification of optimal peptide sequences has been achieved through the screening in solution of SPCLs, each element of which is composed of more than 100,000 nonsupport-bound peptides in equimolar representation, along with an iterative synthesis and screening process. Examples are presented in which an SPCL, composed in total of 52,128,400 acetylated hexa-peptides, is used along with an iterative selection process to precisely identify the antigenic determinant of a peptide recognized by a monoclonal antibody using competitive enzyme-linked immunosorbent assay. This same library was also used to develop highly potent antimicrobial peptides in bacterial growth inhibition assays. A separate non-acetylated SPCL was used to screen and identify high affinity peptide ligands using an opiate radio-receptor binding assay.

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Available from: Clemencia Pinilla, Dec 03, 2014
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    • "As an alternative, positional scanning approach of mixture-based synthetic combinatorial libraries (SCLs) was introduced in 1990s. An SCL is made up of tens of millions of peptide sequences, and positional scanning of such libraries has enabled successful identification of antimicrobial peptides with new or novel properties in a short period of time (Houghten et al., 1991, 1992, 1999; Houghten, 2000). In addition, the SCL approach has been used to improve the activity of existing antimicrobial amphipathic peptides. "
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    ABSTRACT: Antimicrobial peptides (AMPs) are small but effective cationic peptides with variable length. In previous study, four hexapeptides were identified that showed antimicrobial activities against various phytopathogenic bacteria. KCM21, the most effective antimicrobial peptide, was selected for further analysis to understand its modes of action by monitoring inhibitory effects of various cations, time-dependent antimicrobial kinetics, and observing cell disruption by electron microscopy. The effects of KCM21 on Gram-negative strain, Pseudomonas syringae pv. tomato DC3000 and Gram-positive strain, Clavibacter michiganensis subsp. michiganensis were compared. Treatment with divalent cations such as Ca(2+) and Mg(2+) inhibited the bactericidal activities of KCM21 significantly against P. syringae pv. tomato DC3000. The bactericidal kinetic study showed that KCM21 killed both bacteria rapidly and the process was faster against C. michiganensis subsp. michiganensis. The electron microscopic analysis revealed that KCM21 induced the formation of micelles and blebs on the surface of P. syringae pv. tomato DC3000 cells, while it caused cell rupture against C. michiganensis subsp. michiganensis cells. The outer membrane alteration and higher sensitivity to Ca(2+) suggest that KCM21 interact with the outer membrane of P. syringae pv. tomato DC3000 cells during the process of killing, but not with C. michiganensis subsp. michiganensis cells that lack outer membrane. Considering that both strains had similar sensitivity to KCM21 in LB medium, outer membrane could not be the main target of KCM21, instead common compartments such as cytoplasmic membrane or internal macromolecules might be a possible target(s) of KCM21.
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    ABSTRACT: Thesis (Ph. D.)--Massachusetts Institute of Technology, Division of Toxicology, 1996. Vita. Includes bibliographical references (leaves 160-188). by Zheng-Huan Chen. Ph.D.
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