Peptide cross-linking to calmodulin: attachment of [Tyr8]substance P.

Ultraviolet irradiation of calmodulin in the presence of calcium results in either the intramolecular cross-linking of Tyr99 and Tyr138 [Malencik, D.A., & Anderson, S.R. (1986) Biochemistry 25, 709] or, when [Tyr8]substance P is bound, the generation of peptide-calmodulin adducts. The latter consist of two chromatographically distinct fractions, one of which was purified to homogeneity with phenylagarose, DEAE-Sepharose, and reverse-phase chromatography. Chemical characterization shows that the purified conjugate contains 1 mol/mol of peptide covalently attached to Tyr138 of calmodulin. The fluorescence intensity and anisotropy of the dityrosine moiety demonstrate that this novel derivative undergoes interactions with calcium, smooth muscle myosin light chain kinase, and phenylagarose which are similar to those of unmodified calmodulin.