S-100 protein binds to annexin II and p11, the heavy and light chains of calpactin I.
S-100 protein, a dimeric, Ca(2+)-binding protein of the EF-hand type, interacts with annexin II (p36, the heavy chain of the cytoskeletal protein complex, calpactin I), with p11 (the light and regulatory chain of calpactin I) and with the hetero-tetramer annexin II2-p11(2) (calpactin I) in a Ca(2+)-regulated way, but not with annexins I, V and VI. The interaction of S-100 protein with the above proteins was investigated by fluorescence spectroscopy using acrylodan-S-100 protein and acrylodan-annexin II and by cross-linking experiments using the bifunctional cross-linker disuccinimidyl suberate (DSS). S-100 protein binds with the highest affinity to annexin II (Kd approx. 0.4 microM) and with the lowest affinity to calpactin I (Kd approx. 10 microM), with a constant stoichiometry of about 2 mol of protein/S-100 dimer. Thus, S-100 protein could substitute for p11 in regulating the activities of annexin II in cells which do not express p11 and/or act synergistically with p11 in cells expressing both p11 and S-100. The binding of S-100 protein to p11 could reflect the natural tendency of S-100 subunits and p11 to dimerize. Chimeric p11-S-100 alpha and p11-S-100-beta proteins could therefore form in a Ca(2+)-regulated way. The interaction of S-100 protein with calpactin I appears of doubtful physiological importance, because of the low binding affinity, of the small extent of fluorescence changes induced by calpactin I in acrylodan-S-100 protein and of lack of DSS-induced complex formation between the two protein species.
Bianchi
R
R
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Italy.
Pula
G
G
Ceccarelli
P
P
Giambanco
I
I
Donato
R
R
D015081:2-Naphthylamine / Q000031:analogs & derivatives; D000818:Animals; D017306:Annexin A2; D017302:Annexins / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D005456:Fluorescent Dyes; D010455:Peptides / Q000378:metabolism*; D009418:S100 Proteins / Q000378:metabolism*; D000099248:S100 Calcium Binding Protein A10
D015081:2-Naphthylamine; D017306:Annexin A2; D017302:Annexins; D002118:Calcium; D005456:Fluorescent Dyes; D010455:Peptides; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10; C038518:acrylodan