Properties of the reaction of cis-dichlorodiammineplatinum(II) with metallothionein

• Published in: The Journal of biological chemistry 1992-08, Vol.267 (23), p.16121-16128
• Main Authors: A Pattanaik, G Bachowski, J Laib, D Lemkuil, C F Shaw, 3rd, D H Petering, A Hitchcock, L Saryan
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Chlorides - pharmacology; Cisplatin - chemistry; Hydrogen-Ion Concentration; Kinetics; Liver - drug effects; Liver - metabolism; Medical sciences; Metallothionein - biosynthesis; Metallothionein - chemistry; Metallothionein - isolation & purification; Other treatments; Protein Binding; Rats; Rats, Inbred Strains; Spectrophotometry; Treatment. General aspects; Tumors; Zinc - pharmacology; Zinc Compounds
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)41975-8

Properties of the reactions of cis-dichlorodiammine Pt(II) and related complexes with zinc metallothionein or apometallothionein have been investigated. During these reactions, platinum binds stoichiometrically to protein sulfhydryl groups and zinc, if present, is displaced. The ammine ligands are also lost in the process, suggesting that Pt(II) has tetrathiolate coordination in metallothionein. This conclusion is supported by extended x-ray absorption-fine structure studies which indicate that there are 4 sulfurs in the first coordination sphere of the platinum centers. The product contains 10 +/- 2 Pt(II) per mol of protein and migrates over Sephadex G-75 as a structure of similar size to zinc metallothionein. The kinetics of reaction are biphasic as monitored by the formation of Pt-thiolate bonds or by the release of zinc from the protein. Both methods yield identical rate laws for the reaction. The first step is independent of Pt(II) concentration but involves the binding of as many as four platinum atoms to the protein with little Pt-sulfhydryl bond formation and without much loss of zinc. The second rate process is first order in both zinc or sulfhydryl binding sites and Pt(II). Neither kinetic step is sensitive to the chloride ion concentration over the range 0-0.5 M. However, the reaction is sensitive to pH between 5.5 and 8.0. trans-Dichlorodiammineplatinum(II) reacts with zinc metallothionein with similar kinetics.