Characterization of a discontinuous epitope on annexin II by site‐directed mutagenesis

FEBS Letters

Weber, Klaus¹  Gerke, Volker¹  Thiel, Carsten¹ 

[1] Max Planck Institute for Biophysical Chemistry, P.O. Box 2841, D-3400 Göttingen, Germany

关键词: Ca2+/phospholipid-binding protein;    Calpactin;    Lipocortin;    Tyrosine phosphorylation;    cDNA;    complementary DNA;    DTT;    dithiothreitol;    ELISA;    enzyme-linked immunosorbent assay;    EGTA;    [ethylenebis(oxyethylenenitrilo)] tetraacetic acid;    PAGE;    polyacrylamide gel electrophoresis;    pp6Ov-src;    tyrosine kinase encoded by the viral src gene;    RT;    room temperature;    SDS;    sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(91)80724-H

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recombinant annexin II mutants were generated to identify amino acids involved in the formation of the discontinuous epitope of the monoclonal antibody H28. Analysis of the various mutant proteins by immunoblotting and enzyme-linked immunosorbent assay revealed that residues Lys²⁷, Arg⁶², Glu⁶⁵, and Arg⁶⁷ are indispensable for H28 reactivity. Residues in equivalent positions are also in close proximity in the recently determined X-ray structure of annexin V, a different member of the same family of Ca²⁺/lipid-binding proteins. Thus annexins II and V show a similar three-dimensional folding in this region of the molecule. Consequently, the Ca²⁺ binding sites and the residues phosphorylated by pp6O^src (Tyr²³) and protein kinase C (Ser²⁵) most likely reside on opposite sides of the annexin II molecule.

【 授权许可】

Unknown