Springer Protocols: Abstract: Application of Topologically Constrained Mini-Proteins as Ligands, Substrates, and Inhibitors

Application of Topologically Constrained Mini-Proteins as Ligands, Substrates, and Inhibitors

By: Janelle L. Lauer-Fields², Dmitriy Minond³, Keith Brew², Gregg B. Fields²

Abstract

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Protein–protein interactions are governed by a variety of structural features. The sequence specificities of such interactions are usually easier to establish than the “topological specificities,” whereby interactions may be classified based on recognition of distinct three-dimensional structural motifs. Approaches to explore topological specificities have been based primarily on assembly of mini-proteins with well defined secondary, tertiary, and/or quarternary structures. The present chapter focuses on three approaches for constructing topologically well defined mini-proteins: template-assembled synthetic proteins (TASPs), disulfide-stabilized structures, and peptide-amphiphiles (PAs). Specific examples are given for applying each approach to explore topologically-dependent protein–protein interactions. TASPs are utilized to identify a metastatic melanoma receptor that binds to the α1(IV)1263–1277 region of basement membrane (type IV) collagen. A disulfide-stabilized structure incorporating a sarafotoxin (SRT) 6b model was examined as a matrix metalloproteinase (MMP)-3 inhibitor. PAs were developed as (a) fluorogenic triple-helical or polyPro II substrates for MMPs and aggrecanase members of the a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) family and (b) glycosylated and nonglycosylated ligands for metastatic melanoma cells. Topologically constrained mini-proteins have proved to be quite versatile, helping to define critical primary, secondary, and tertiary structural elements that modulate enzyme and receptor functions.

Affiliation(s): (2) College of Biomedical Sciences and Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, FL
(3) Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, FL

Book Title: Peptide Characterization and Application Protocols

Series: Methods in Molecular Biology | Volume: 386 | Pub. Date: May-11-2007 | Page Range: 125-166 | DOI: 10.1007/978-1-59745-430-8_5

Subject: Protein Science

Key Words: Triple-helix - peptide-amphiphile - matrix metalloproteinase - collagen - integrin

References

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