title: Covalent sequestration of the nitrogen mustard mechlorethamine by metallothionein.
abstract: The research reported here demonstrates covalent binding to the metal-binding protein metallothionein (MT) by the therapeutic nitrogen mustard mechlorethamine. The most surprising aspect of this interaction is the selectivity of the alkylating agent for specific residues of MT. A combination of MS and proteolytic and enzymatic methods was used to deduce specific locations of mechlorethamine alkylation. These experiments indicated that alkylation occurs predominantly in the carboxyl domain of MT, with one molecule of mechlorethamine covalently cross-linking two cysteine residues. Electrospray MS revealed the retention of all seven metal ions in the cross-linked MT/mechlorethamine adducts, highlighting the uniqueness of this protein. Computerized docking experiments supported the hypothesis that selective binding precedes selective alkylation, and the structure of the drug indicates the minimal structural requirements for this binding. These results support the idea that MT overexpressed in tumor cells contributes to the inactivation of anticancer drugs.
authors.affiliation: Structural Biochemistry Center, University of Maryland Baltimore County, College Park, 20742, USA.
authors.forename: M
authors.initials: M
authors.lastname: Antoine
authors.forename: D
authors.initials: D
authors.lastname: Fabris
authors.forename: C
authors.initials: C
authors.lastname: Fenselau
mesh_terms: D000477:Alkylating Agents / Q000493:pharmacokinetics*; D000478:Alkylation; D000818:Animals; D002268:Carboxypeptidases / Q000378:metabolism; D003198:Computer Simulation; D006868:Hydrolysis; D066298:In Vitro Techniques; D008466:Mechlorethamine / Q000493:pharmacokinetics*; D008668:Metallothionein / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D011817:Rabbits; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin
chemical_list: D000477:Alkylating Agents; D008466:Mechlorethamine; D008668:Metallothionein; D002268:Carboxypeptidases; D014357:Trypsin