Min K L, Steghens J P, Henry R, Doutheau A, Collombel C
Laboratoire de Biochimie C, Hôpital Edouard Herriot, 5 place d'Arsonval, F-69437 Lyon Cedex 03, France.
Biochim Biophys Acta. 1998 Sep 8;1387(1-2):80-8. doi: 10.1016/s0167-4838(98)00107-1.
A new photoaffinity probe with a benzophenone group, N-dibenzylphospho-N'-(4-benzoyl)-benzylguanidine (BzPG), has been synthesized on the basis on our previously described creatine kinase bisubstrate analog. BzPG is also a bisubstrate type analog whose photoinsertion is inhibited by the natural substrates of creatine kinase. When rabbit CK-MM is irradiated in the presence of BzPG then cleaved by CNBr, one labeled peptide can be purified by reverse phase HPLC and sequenced. This sequence of 31 amino acids (Ala30-Val60) contains a region which could be responsible for isoenzyme selectivity and another one just preceding the 11 amino acid peptide (Asp61-Thr70) very recently described as a putative creatine binding site. This second peptide was deduced from the comparison of 18 amino acid sequence alignments. We proposed the creatine binding site to be essentially a peptide from Lys39 to Val71.
