Peptide‐specific antibodies localize the major lipid binding sites of talin dimers to oppositely arranged N‐terminal 47 kDa subdomains

FEBS Letters

Goldmann, Wolfgang H.²  Isenberg, Gerhard¹ 

[1] Biophysics Department E-22, Technical University of Munich, James-Franck Strasse, D-85747 Garching, Germany;Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA

关键词: Electron microscopy;    Lipid binding site;    Peptide-specific antibody;    Talin;    DMPC;    1;    2-dimyristoyl phosphatidylcholine;    DMPG;    1;    2-dimyristoyl phosphatidylglycerol;    DPPC;    1;    2-dipalmitoyl-sn-glycero-3-phosphatidylcholine;   

DOI  :  10.1016/S0014-5793(98)00336-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using ultrastructural analysis and labeling with polyclonal antibodies that recognize peptide sequences specific for phospholipid binding, we mapped the functional domain structure of intact platelet talin and its proteolytic fragments. The talin dimer, which is crucial for actin and lipid binding, is built of a backbone containing the 200 kDa rod portions, at both ends of which a 47 kDa globular domain is attached. Peptide-specific polyclonal antibodies were raised against three potential lipid binding sequences residing within the N-terminal 47 kDa domain (i.e. S19, amino acids 21–39; H18, amino acids 287–304; and H17, amino acids 385–406). Antibodies H17 and H18 localize these lipid binding sequences within the N-terminal 47 kDa globular talin subdomains opposed at the outer 200 kDa rod domains within talin dimers. Hence, we conclude that in its dimeric form, which is used in actin and lipid binding, talin is a dumbbell-shaped molecule built of two antiparallel subunits.

【 授权许可】

Unknown