INSULIN-LIKE GROWTH-FACTOR (IGF)-II BINDING TO IGF-BINDING PROTEINS AND IGF RECEPTORS IS MODIFIED BY DELETION OF THE N-TERMINAL HEXAPEPTIDEOR SUBSTITUTION OF ARGININE FOR GLUTAMATE-6 IN IGF-II

Recombinant insulin-like growth factor-II (IGF-II) and two structural analogues, des(1-6)IGF-II and [Arg6]-IGF-II, were produced to investig ate the role of N-terminal residues in binding to IGF-binding proteins (IGFBPs) and hence the biological properties of the modified peptides . The growth factors were modelled on two previously characterized var iants of IGF-I, des(1-3)IGF-I and [Arg3]-IGF-I, which both show substa ntially decreased binding to IGFBPs and were expressed as fusion prote ins in Eschericia coli. The biological activities of the corresponding analogues of IGF-I and IGF-II were compared in rat L6 myoblasts and H 35B hepatoma cells. In the L6-myoblast protein-synthesis assay, the IG F-II analogues, des(1-6)IGF-II and [Arg6]-IGF-II, were slightly more p otent than IGF-II but about 10-fold less potent than IGF-I and 100-fol d less potent than the respective IGF-I analogues, des(1-3)IGF-I and [ Arg3]-IGF-I. In H35 hepatoma cells the anabolic response measured was the inhibition of protein breakdown, and the potency order was insulin much greater than [Arg3]-IGF-I > des(I-3)IGF-I > [Arg6]-IGF-II > des( 1-6)IGF-II > IGF-I > IGF-II. Binding of the IGFs and their analogues t o the type 1 IGF receptor in L6 myoblasts and to the insulin receptor in H35 hepatoma cells did not fully explain the observed anabolic pote ncy differences. Moreover, binding of all four analogues to the IGFBPs secreted by L6 myoblasts and H35B hepatoma cells was greatly decrease d compared with the parent IGF. We conclude that the observed anabolic response to each IGF was determined by their relative binding to the competing cell receptor and IGFBP binding sites present.