Application of topologically constrained mini-proteins as ligands, substrates, and inhibitors.
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 alpha1(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.
Lauer-Fields
Janelle L
JL
College of Biomedical Sciences and Department of Chemistry & Biochemistry, Florida Atlantic University, Boca Raton, USA.
Minond
Dmitriy
D
Brew
Keith
K
Fields
Gregg B
GB
D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004220:Disulfides / Q000737:chemistry; D004355:Drug Stability; D006801:Humans; D008024:Ligands; D061965:Matrix Metalloproteinase Inhibitors; D008958:Models, Molecular; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D019714:Tissue Inhibitor of Metalloproteinases / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D014757:Viper Venoms / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology
D004220:Disulfides; D008024:Ligands; D061965:Matrix Metalloproteinase Inhibitors; D011506:Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D014757:Viper Venoms; C057354:sarafotoxins s6