Development of a high-throughput screening system for the compounds that inhibit collagen-protein interactions.
Collagen-binding proteins (CBPs) play important roles in various physiological events. Some CBPs are regarded as targets for drug development; for example, platelet glycoprotein VI (GPVI) and heat shock protein 47 (HSP47) are promising targets for the development of novel antiplatelet and antifibrotic drugs, respectively. However, no systematic screening method to search compounds that inhibit collagen-CBP interactions have been developed, and only a few CBP inhibitors have been reported to date. In this study, a facile turbidimetric multiwell plate assay was developed to evaluate inhibitors of CBPs. The assay is based on the finding that CBPs retard spontaneous collagen fibril formation in vitro and that fibril formation is restored in the presence of compounds that interfere with the collagen-CBP interactions. Using the same platform, the assay was performed in various combinations of fibril-forming collagen types and CBPs. This homogeneous assay is simple, convenient, and suitable as an automated high-throughput screening system.
Okano-Kosugi
Hitomi
H
Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata 956-8603, Japan.
Matsushita
Osamu
O
Asada
Shinichi
S
Herr
Andrew B
AB
Kitagawa
Kouki
K
Koide
Takaki
T
D000818:Animals; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005240:Feasibility Studies; D050958:HSP47 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D009391:Nephelometry and Turbidimetry; D011485:Protein Binding / Q000187:drug effects; D013552:Swine
D050958:HSP47 Heat-Shock Proteins; D003094:Collagen