Comparative Structural Studies of the Active Site of ATP: Guanidine Phosphotransferases

Abstract

The active thiol group of lombricine kinase from Lumbricus terrestris muscle was labelled with N ‐ethyl‐[1‐ 14 C]maleimide. The resulting inactivated N ‐ethyl‐[1‐ 14 C]succinimido enzyme was then subjected to tryptic hydrolysis. The peptide containing the labelled essential thiol group was isolated and found to contain: Leu‐Gly‐Tyr‐Ile‐Thr‐[ 14 C]Cys‐Pro‐Gly‐Ser‐Asn‐Leu‐Gly‐Thr‐Leu‐Arg. The amino acid sequence around this thiol group was very similar with that of homologous ATP: guanidine phosphotransferases previously studied, arginine kinase from Homarus vulgaris muscle, creatine kinases from ox brain and ox muscle and from rabbit muscle. In addition among the other enzymes of this group, lombricine kinase is of special interest since it is the only dimeric enzyme of molecular weight ≃ 80000 which possesses only one essential thiol group and one nucleotide binding site per two subunits.