title: Application of mercury cold vapor atomic fluorescence spectrometry to the characterization of mercury-accessible -SH groups in native proteins.
abstract: A new analytical approach has been applied to the determination and characterization of mercury-accessible -SH groups in pure native protein samples (ovalbumin, hemoglobin, glyceraldehyde-3-phosphate dehydrogenase, aldolase, pyruvate kinase, hexokinase, lactate dehydrogenase, alcohol dehydrogenase, creatine phosphokinase, lysozyme, and cytochrome c). The method is based on the selective reduction of Hg(II) in the presence of Hg(II)-thiol complexes with alkaline sodium tetrahydroborate, to give Hg(0) in a continuous flow reaction system coupled with atomic fluorescence spectrometric (AFS) detection. The method is fast and specific and allows one to work with nanomole amounts of a single protein without any preliminary incubation and without any separation of Hg(II) from thiol-complexed mercury. The meaning of the results obtained in the determination of the accessible -SH groups in native proteins by using chemical probes is discussed.
authors.affiliation: Institute of Instrumental Analytical Chemistry, Department of Chemistry and Industrial Chemistry, CNR, Via Risorgimento 35, Pisa, 56126, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Bramanti
authors.forename: A
authors.initials: A
authors.lastname: D'Ulivo
authors.forename: L
authors.initials: L
authors.lastname: Lampugnani
authors.forename: R
authors.initials: R
authors.lastname: Zamboni
authors.forename: G
authors.initials: G
authors.lastname: Raspi
mesh_terms: D000818:Animals; D003080:Cold Temperature; D006801:Humans; D017669:Mercury Compounds; D011506:Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013438:Sulfhydryl Compounds; D014835:Volatilization
chemical_list: D017669:Mercury Compounds; D011506:Proteins; D013438:Sulfhydryl Compounds