title: Role of Tyr84 in controlling the reactivity of Cys34 of human albumin.
abstract: Cys34 in domain I of the three-domain serum protein albumin is the binding site for a wide variety of biologically and clinically important small molecules, provides antioxidant activity, and constitutes the largest portion of free thiol in blood. Analysis of X-ray structures of albumin reveals that the loop containing Tyr84 occurs in multiple conformations. In structures where the loop is well defined, there appears to be an H-bond between the OH of Tyr84 and the sulfur of Cys34. We show that the reaction of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) with Tyr84Phe mutant albumin is approximately four times faster than with the wild-type protein between pH 6 and pH 8. In contrast, the His39Leu mutant reacts with DTNB more slowly than the wild-type protein at pH < 8, but at a similar rate at pH 8. Above pH 8 there is a dramatic increase in reactivity for the Tyr84Phe mutant. We also report (1)H NMR studies of disulfide interchange reactions with cysteine. The tethering of the two loops containing Tyr84 and Cys34 not only appears to control the redox potential and accessibility of Cys34, but also triggers the transmission of information about the state of Cys34 throughout domain I, and to the domainI/II interface.
authors.affiliation: School of Chemistry, University of Edinburgh, West Mains Road, Edinburgh, EH9 3JJ, UK.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Stewart
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Blindauer
authors.forename: Stephen
authors.initials: S
authors.lastname: Berezenko
authors.forename: Darrell
authors.initials: D
authors.lastname: Sleep
authors.forename: David
authors.initials: D
authors.lastname: Tooth
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Sadler
mesh_terms: D001665:Binding Sites; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D012709:Serum Albumin