HOW THE ANTI-(METAL CHELATE) ANTIBODY CHA255 IS SPECIFIC FOR THE METAL-ION OF ITS ANTIGEN - X-RAY STRUCTURES FOR 2 FAB' HAPTEN COMPLEXES WITH DIFFERENT METALS IN THE CHELATE

Citation
Ra. Love et al., HOW THE ANTI-(METAL CHELATE) ANTIBODY CHA255 IS SPECIFIC FOR THE METAL-ION OF ITS ANTIGEN - X-RAY STRUCTURES FOR 2 FAB' HAPTEN COMPLEXES WITH DIFFERENT METALS IN THE CHELATE, Biochemistry, 32(41), 1993, pp. 10950-10959
Citations number
54
Categorie Soggetti
Biology
Journal title
ISSN journal
00062960
Volume
32
Issue
41
Year of publication
1993
Pages
10950 - 10959
Database
ISI
SICI code
0006-2960(1993)32:41<10950:HTACAC>2.0.ZU;2-Z
Abstract
Antibodies with bound metal-chelate haptens provide new means for expl oiting the diverse properties of metallic elements. The murine monoclo nal antibody CHA255 (IgG1lambda) binds the metal-chelate hapten I)-4-[ N'-(2-hydroxyethyl)thioureido]-L-benzyl-EDTA (designated In-EOTUBE) wi th high affinity (K(a) = 1.1 x 10(10) M-1). Antibody binding, is highl y specific for the indium chelate; the affinity decreases as much as 1 0(4) with other metals, even those, having ionic radii close to indium . To better understand this selectivity, the crystal structure of the antigen-binding fragment (Fab') of CHA255 complexed with its hapten, I n(III)-EOTUBE, was determined by molecular replacement and refined at 2.2-angstrom resolution. The structure of CHA255 Fab' complexed with F e(III)-EOTUBE was also determined and refined at 2.8-angstrom resoluti on. In both structures, the hapten's EDTA moiety is half-buried near t he center of the complementarity-determining regions (CDR's). Five of the six CDR's on the Fab' interact with the hapten through protein sid e-chain atoms (but not main-chain atoms). A novel feature of the In-EO TUBE/Fab' complex is coordination of the indium by Nepsilon of one his tidine from the heavy chain's third CDR (distance = 2.4 angstrom). The histidine coordination is not observed in the Fe-EOTUBE/Fab' complex, due mainly to a slightly different hapten conformation that reduces m etal accessibility; this may partially explain the 20-fold lower affin ity of CHA255 for iron hapten. An unexpected feature of the Fab' overa ll is an elbow angle of 193-degrees (the angle between the pseudodyad axes of the Fab's constant and variable domains).