The amino‐terminal fragment of gelsolin is cross‐linked to Cys‐374 of actin in the EGTA‐resistant actin‐gelsolin complex

FEBS Letters

Doi, Yukio¹  Kanatani, Yumiko¹  Kim, Fimi² 

[1] Department of Food Science, Kyoto Women's University, Higashiyama-ku, Kyoto 605, Japan;Protein Engineering Research Institute, 6-2-3 Furuedai, Suita-shi, Osaka 565, Japan

关键词: Gelsolin;    Actin;    Actin binding protein;    BPM;    benzophenone-4-maleimide;    EDC;    1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide;    EGTA;    [ethylenebis(oxyethelenenitrilo)]tetraacetic acid;    F-actin;    filamentous actin;    G-actin;    globular actin;    NBD-Cl;    7-chloro-4-nitrobenzeno-2-oxa-1;    3-diazole chloride;    NBD;    7-nitrobenz-2-oxa-1;    3-diazole-4-yl;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(92)80218-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

It has been shown that the EGTA-resistant actin, one of the two actin molecules associated to gelsolin, can be predominantly cross-linked to gelsolin by benzophenone-4-maleimide (BPM), a photoaffinity-labeling reagent, which was conjugated to Cys-374 of actin prior to cross-linking (Doi, Y., Banba, M. and Vertut-Doï, A. (1991) Biochemistry 30, 5769–5777). When a chymotryptic digest of gelsolin containing the amino-terminal 15-kDa fragment was mixed with BPM-actin (42 kDa) and irradiated for cross-linking, a band of 58 kDa appeared on SDS-PAGE which was shown to contain actin molecule by using fluorescently labeled actin. The amino-terminal sequence of the 58-kDa complex was identical to that of gelsolin, confirming that the amino-terminal segment (residues 1–133) of pig plasma gelsolin lies closely to Cys-374 of actin in the EGTA-resistant complex.

【 授权许可】

Unknown