Noncovalent associations of glutathione S-transferase and ligands: A study using electrospray quadrupole/time-of-flight mass spectrometry
- Masaki IshigaiAffiliated withMichael Barber Centre for Mass Spectrometry, UMISTFuji Gotenba Research Lab, Chugai Pharmaceutical Co. Ltd.
- , James I. LangridgeAffiliated withMicromass UK Ltd.
- , Robert S. BordoliAffiliated withMicromass UK Ltd.
- , Simon J. GaskellAffiliated withMichael Barber Centre for Mass Spectrometry, UMIST Email author
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Human glutathione S-transferase A1-1 was observed predominantly as dimeric ions (51 kDa) during electrospray mass spectrometric analysis from aqueous solution at pH 7.4, in keeping with the known dimeric structure in solution. When analyses were performed on solutions of the enzyme containing glutathione (GSH), noncovalent adducts of protein dimer and one or two ligand molecules were observed; each mass increment, which exceeded the mass of GSH alone, was provisionally interpreted to indicate concomitant association of two water molecules per bound GSH. Noncovalent adducts of ligand and protein dimer were similarly observed for oxidized glutathione and for two glutathione inhibitors, both incorporating substituted thiol structures. In these instances, the mass increments exactly matched the ligand masses, suggesting that the apparent concomitant binding of water was associated with the presence in the ligand of a free thiol group. Collisionally activated decomposition during tandem mass spectrometry analyses of noncovalent adducts incorporating protein dimer and ligands yielded initially the denuded dimer; at higher collision energies the monomer and a protein fragment were formed.
- Noncovalent associations of glutathione S-transferase and ligands: A study using electrospray quadrupole/time-of-flight mass spectrometry
Journal of the American Society for Mass Spectrometry
Volume 11, Issue 7 , pp 606-614
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- 1. Michael Barber Centre for Mass Spectrometry, UMIST, M60 1QD, Manchester, UK
- 2. Fuji Gotenba Research Lab, Chugai Pharmaceutical Co. Ltd., 135, Komakado 1-chome, Gotenba-shi, 412-8513, Shizuoka, Japan
- 3. Micromass UK Ltd., Manchester, UK