Preparation and characteristics of Ca(2+)-dependent monoclonal antibodies to recoverin.
Thirty-four primary hybridoma clones were prepared which expressed monoclonal antibodies to the Ca(2+)-binding protein recoverin. Among the resulting monoclonal antibodies, two Ca(2+)-dependent clones (mAb3 and mAb19) recognizing recoverin were detected by solid-phase immunoenzyme assay. In the presence of Ca(2+), antibodies of the mAb3 and mAb19 clones bound to recoverin several times better than in the absence of Ca(2+). The mAb3 and mAb19 antibodies recognized epitopes located inside the sequences Pro61-Met91 and Pro57-Tyr64 of the recoverin molecule, respectively. The possible mechanism of the Ca(2+)-dependent recognition of recoverin by the prepared monoclonal antibodies is discussed.
Tikhomirova
N K
NK
Department of Cell Signal Systems, Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow 119992, Russia.
Goncharskaya
M A
MA
Senin
I I
II
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015151:Immunoblotting; D007340:Insulinoma; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D051603:Recoverin; D013154:Spleen / Q000166:cytology
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008074:Lipoproteins; C496732:Rcvrn protein, mouse; D051603:Recoverin; D002118:Calcium