PHOTOCONTROL OF MYOSIN V ATPASE ACTIVITY USING CALMODULIN MODIFIED WITH PHOTOCHROMIC COMPOUND
PHOTOCONTROL OF MYOSIN V ATPASE ACTIVITY USING CALMODULIN MODIFIED WITH PHOTOCHROMIC COMPOUND
复制标题
使用光致变色化合物修饰的钙调蛋白对肌球蛋白 V ATP酶活性的光控制
DOI：
10.1016/j.bpj.2010.12.2854
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发表时间：
2011
影响因子：
3.4
通讯作者：
Mitsuo Ikebe and Shinsaku Maruta
中科院分区：
文献类型：
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作者：
Hideki Shishido;Mitsuo Ikebe and Shinsaku Maruta
钙调素（Calmodulin，CaM）是一种重要的钙离子结合蛋白，参与多种细胞调节过程. CaM在与Ca2+结合后发生构象变化，这使得它能够与特定的蛋白质结合以产生特定的反应。例如，Ca2 +/CaM调节肌动蛋白激活的肌球蛋白V的ATP酶活性。肌球蛋白V是一种进行性马达，在各种细胞中具有作为细胞器转运体的作用。肌球蛋白V的颈区带有六个IQ基序，它们是CaM或CaM样轻链的结合位点，我们利用光致变色化合物N-（4-phenylazophenyl）maleimide（PAM）可逆地发生顺反异构化，成功地调控了CaM与其靶肽的结合。为了将PAM掺入到特定位点，制备了在功能区中具有反应性半胱氨酸残基的CaM突变体。PAM被化学计量地掺入这些突变体中的半胱氨酸残基中。PAM CaM突变体N60 C、D64 C和M124 C与CaM靶肽的结合在适当的Ca2+浓度下受到UV-VIS光照射的可逆光控制。
Calmodulin (CaM) is a physiologically important Ca 2+-binding protein that participates in numerous cellular regulatory processes. CaM undergoes a conformational change upon binding to Ca 2+, which enables it to bind to specific proteins for specific responses. For example, Ca 2+/CaM regulates actin-activated ATPase activity of myosin V. Myosin V is a processive motor that has a role as an organelle transporter in various cells. The neck domain of myosin V carries six IQ motifs, which act as a binding site for CaM or CaM-like light chains.Previously, we succeeded to photocontrol 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 incorporate PAM into specific site, CaM mutants that have a reactive cysteine residue in the functional region were prepared. PAM was stoichiometrically incorporated into the cysteine residues in these mutants. The binding of the PAM-CaM mutants, N60C, D64C, and M124C, to CaM target peptide was reversibly photocontrolled upon UV-VIS light irradiation at appropriate Ca 2+ concentrations.