1995 Fiscal Year Final Research Report Summary

Construction and Characterization of Calcium Responsive Protein Assembling

Research Project

Project/Area Number	06805071
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	生物・生体工学
Research Institution	Tokyo Institute of Technology
Principal Investigator	KOBATAKE Eiry Tokyo Institute of Technology, Department of Bioengineering, Associate Professor, 生命理工学部, 講師 (00225484)
Project Period (FY)	1994 – 1995
Keywords	Molecular Assembling / Calmodulin / Fusion Protein / LB membrane / Calcium Ion / Glutathione S-Transferase
Research Abstract	In this study, some methods were performed for assembling protein molecules. Calmodulin, a calcium ion responsive protein, was conjugated with lipid and subsequently assembled into a monolayr using LB method. The lipid-conjugated calmodulin retains its calcium sensitivity, determined from the changes in the area-pressure isotherm of the monolayer obtained at the air-water interface. To assemble calmodulin on a solid-phase matrix, a fusion protein consisting of calmodulin and glutathione S-transferase (GST) was produced by gene fusion. The protein had the modulating activity of calmodulin and binding capability to glutathione of GST.Furthermore, calmodulin could be immobilized on a solid-phase matrix through the use of GST moiety which modulating activity was retained. The activity of phosphodiesterase, which is a calmodulin dependent enzyme, was controlled by the immobilized calmodulin with calcium ion, and repeated use of calmodulin was demonstrated.

(5 results)