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Characterization of a discontinuous epitope on annexin II by site‐directed mutagenesis
Author(s) -
Thiel Carsten,
Weber Klaus,
Gerke Volker
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80724-h
Subject(s) - epitope , annexin , microbiology and biotechnology , mutagenesis , chemistry , monoclonal antibody , site directed mutagenesis , mutant , recombinant dna , biochemistry , binding site , amino acid , biology , antibody , in vitro , gene , immunology
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 Lys 27 , Arg 62 , Glu 65 , and Arg 67 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 Ca 2+ /lipid‐binding proteins. Thus annexins II and V show a similar three‐dimensional folding in this region of the molecule. Consequently, the Ca 2+ binding sites and the residues phosphorylated by pp6O src (Tyr 23 ) and protein kinase C (Ser 25 ) most likely reside on opposite sides of the annexin II molecule.