SYNTHESIS OF COBALAMIN-BIOTIN CONJUGATES THAT VARY IN THE POSITION OFCOBALAMIN COUPLING - EVALUATION OF COBALAMIN DERIVATIVE BINDING TO TRANSCOBALAMIN-II

Citation
Pm. Pathare et al., SYNTHESIS OF COBALAMIN-BIOTIN CONJUGATES THAT VARY IN THE POSITION OFCOBALAMIN COUPLING - EVALUATION OF COBALAMIN DERIVATIVE BINDING TO TRANSCOBALAMIN-II, Bioconjugate chemistry, 7(2), 1996, pp. 217-232
Citations number
72
Categorie Soggetti
Biology,Chemistry,"Biochemical Research Methods
Journal title
ISSN journal
10431802
Volume
7
Issue
2
Year of publication
1996
Pages
217 - 232
Database
ISI
SICI code
1043-1802(1996)7:2<217:SOCCTV>2.0.ZU;2-F
Abstract
Six cobalamin-biotin conjugates have been prepared. The cobalamin-biot in conjugates were prepared to evaluate the effect that the location o f attachment had on the binding with transcobalamin II (TCII), the cob alamin binding protein in plasma, and to evaluate their potential use for in vitro and in vivo applications. This study focused only on the effect of binding with TCII. To decrease the possibility of steric pro blems in binding of the cobalamin conjugates with TCII, and biotin's b inding with streptavidin or avidin, moieties of 11-18 atoms in length were used as linkers. Four biotin conjugates were prepared which were attached to the corrin ring of the cobalamin molecule (on b-, c-, d-, and e-side chains). One conjugate was attached to the 5'-OH of the rib ose moiety, and another conjugate was attached at the cobalt metal (in place of the cyanide moiety of cyanocobalamin). Competitive binding s tudies were conducted where various amounts of the cobalamin-biotin co njugates and their precursor cobalamin derivatives competed with [Co-5 7]cyanocobalamin for binding of recombinant human TCII (rhTCII). Evalu ation of cobalamin derivatives which were conjugated at the 5'-OH of r ibose or the cobalt metal center indicated that conjugation at either of these positions had Little effect on binding with rhTCII. However, conjugates where the attachment was made on the corrin ring substituen ts had a large variation in binding with rhTCII. Conjugates on the e-p ropionamide side chain had little effect (relative affinity was equal to or decreased less than a factor of 3) on binding with rhTCII, conju gates of the b-isomer had decreased binding (relative affinity decreas ed less than a factor of 10), conjugates of the d-propionamide had fur ther decreased binding (relative affinity decreased between 44 and 69 times), and conjugates on the c-acetamide group had poor binding to rh TCII (relative affinity decreased between 295 and 1160 times). The sig nificance of the side chains on the corrin ring in providing specifici ty and high-affinity binding with rhTCII is discussed.