Photocontrol of calmodulin interaction with target peptides using azobenzene derivative.

Abstract:

: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.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Shishido H,Yamada MD,Kondo K,Maruta S

doi

10.1093/jb/mvp107

subject

Has Abstract

pub_date

2009-10-01 00:00:00

pages

581-90

issue

4

eissn

0021-924X

issn

1756-2651

pii

mvp107

journal_volume

146

pub_type

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