Monitoring metal ion flux in reactions of metallothionein and drug-modified metallothionein by electrospray mass spectrometry.
The capabilities of electrospray ionization mass spectrometry are demonstrated for monitoring the flux of metal ions out of and into the metalloprotein rabbit liver metallothionein and, in one example, chlorambucil-alkylated metallothionein. Metal ion transfers may be followed as the reactions proceed in situ to provide kinetic information. More uniquely to this technique, metal ion stoichiometries may be determined for reaction intermediates and products. Partners used in these studies include EDTA, carbonic anhydrase, a zinc-bound hexamer of insulin, and the core domain of bacteriophage T4 gene 32 protein, a binding protein for single-stranded DNA.
Zaia
J
J
Department of Chemistry and Biochemistry and the University of Maryland Cancer Center, University of Maryland Baltimore County, 21250, USA.
Fabris
D
D
Wei
D
D
Karpel
R L
RL
Fenselau
C
C
D000478:Alkylation; D000818:Animals; D017122:Bacteriophage T4; D002256:Carbonic Anhydrases / Q000378:metabolism; D002699:Chlorambucil / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004492:Edetic Acid / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008668:Metallothionein / Q000737:chemistry* / Q000378:metabolism*; D008670:Metals / Q000378:metabolism*; D011817:Rabbits; D014764:Viral Proteins / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
D004268:DNA-Binding Proteins; D007328:Insulin; D008670:Metals; D014764:Viral Proteins; C060035:gp32 protein, Enterobacteria phage T4; D002699:Chlorambucil; D008668:Metallothionein; D004492:Edetic Acid; D002256:Carbonic Anhydrases; D015032:Zinc