Structural requirements of KTS-disintegrins for inhibition of α1β1 integrin

Obtustatin and viperistatin represent the shortest known snake venom monomeric disintegrins. In the present Study, we have produced recombinant full-length wild-type and site-directed mutants of obtustatin to assess the role of the (KTS23)-T-21 tripeptide and C-terminal residues for specific inhibition of the alpha(1)beta(1) integrin. Thr(22) appeared to be the most critical residue for disintegrin activity, whereas Substitution of the flanking lysine or serine residues for alanine resulted in a less pronounced decrease in the anti-alpha(1)beta(1), integrin activity of the disintegrin. The triple mutant A(21)AA(23) was devoid of blocking activity towards alpha(1)beta(1) integrin-mediated cell adhesion. file potency of recombinant KTS disintegrins also depended oil the residue C-terminally adjacent to the active motif. substitution of Leu(24) of wild-type obtustatin for an alanine residue slightly decreased the inhibitory activity of the mutant, whereas an arginine residue in this position enhanced the potency of he mutant over wild-type obtustatin by 6-fold. In addition, the replacements L38V and P40Q may account for it further 25-fold increase in alpha(1)beta(1) inhibitory potency of viperistatin over KTSR-obtustatin.