title: Evidence that the principal CoII-binding site in human serum albumin is not at the N-terminus: implication on the albumin cobalt binding test for detecting myocardial ischemia.
abstract: Human serum albumin (HSA) is the most abundant protein in the blood plasma and is involved in the transport of metal ions. Four metal-binding sites with different specificities have been described in HSA: (i) the N-terminal site provided by Asp1, Ala2, and His3, (ii) the site at the reduced Cys34, (iii) site A, including His67 as a ligand, and (iv) the nonlocalized site B. HSA can bind CoII, and HSA was proposed to be involved in CoII transport. Recently, binding of CoII to HSA has attracted much interest due to the so-called albumin cobalt binding (ACB) test approved by the Food and Drug Administration for evaluation of myocardial ischemia. Although the binding of CoII to HSA is important, the binding of CoII to HSA is not well-characterized. Here the binding of CoII to HSA was studied under anaerobic conditions to prevent CoII oxidation. Electronic absorption, EPR, and NMR spectroscopies indicate three specific and well-separated binding sites for CoII in HSA. CoII ions in all three sites are in a high-spin state and coordinated in a distorted octahedral geometry. Competition experiments with CdII (known to bind to sites A and B) and CuII (known to bind to the N-terminal site) were used to identify the sites of binding of CoII to HSA. They revealed that the first two equivalents of CoII bind to sites A and B. Only the third may be bound to the N-terminal site. The repercussions of these results on the understanding of the ACB test and hence the myocardial ischemia are discussed.
authors.affiliation: Laboratoire de Chimie de Coordination, CNRS UPR 8241, associated with the University of Toulouse III, 205 route de Narbonne, 31077 Toulouse Cedex, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Mothes
authors.forename: Peter
authors.initials: P
authors.lastname: Faller
mesh_terms: D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism; D003035:Cobalt / Q000378:metabolism*; D003300:Copper / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D017202:Myocardial Ischemia / Q000175:diagnosis*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D012709:Serum Albumin; D002104:Cadmium; D003035:Cobalt; D003300:Copper