| 19605460 |
Shishido H, Yamada MD, Kondo K, Maruta S: Photocontrol of calmodulin interaction with target peptides using azobenzene derivative. J Biochem. 2009 Oct;146(4):581-90. Epub 2009 Jul 15.
Calmodulin (CaM), a physiologically important Ca (2+)-binding protein, participates in numerous cellular regulatory processes. It is dumbbell shaped and contains two globular domains connected by a short alpha-helix. Each of the globular domains has two Ca (2+)-binding sites, the EF hands. CaM undergoes a conformational change upon binding to Ca (2+), which enables it to bind to specific proteins for specific responses. Here, we successfully photocontrolled CaM binding to its target peptide using the photochromic compound N-(4-phenylazophenyl) maleimide (PAM), which reversibly undergoes cis-trans isomerization upon ultraviolet (UV) and visible (VIS) light irradiation. In order to specifically incorporate PAM, CaM mutants having reactive cysteine residues in the functional region were prepared; PAM was stoichiometrically incorporated into the cysteine residues in these mutants. Further, we prepared the target peptide, M13, fused with yellow fluorescent protein (YFP) to monitor the CaM-M13 peptide interaction. The binding of the PAM-CaM mutants, N60C, D64C and M124C, to M13-YFP was reversibly photocontrolled upon UV-VIS light irradiation at appropriate Ca (2+) concentrations. |
82(1,1,1,2) |