Heavy metal ion interactions with Callinectes sapidus hemocyanin: structural and functional changes induced by a variety of heavy metal ions.

Journal Article (Journal Article)

Hemocyanins are oligomeric proteins that reversibly bind oxygen. The oxygen binding site is a binuclear copper center bound to the protein by amino acid side chains. The hemocyanin of the blue crab, Callinectes sapidus, occurs in vivo as a mixture of 25S dodecamers and 16S hexamers, whose oxygen binding properties are identical. Four heavy metals have been used as probes of structure and function in this hemocyanin system. Divalent cations of cadmium, copper, mercury, and zinc induced an indefinite self-association of the hemocyanin molecule. These higher ordered association states can be dissociated by ethylenediaminetetraacetic acid. Callinectes oxyhemocyanin possesses at least three mercury binding sites: (1) a sulfhydryl group which forms a mercaptide bond with a single mercuric ion, (2) a tryptophanyl side chain which forms a noncovalent 1:1 complex with mercuric ions with an association constant of 5.7 X 10(15) M-1, and (3) lower affinity site(s) involved in the self-association process also observed with cadmium, copper, and zinc. Sites 1 and 2 are most likely also involved in the binding of cadmium. Upon removal of oxygen from the active site of hemocyanin, an additional binding site becomes available for the reaction with mercury. Binding of mercury to this site leads to loss of one of the coppers from the binuclear oxygen binding site. Both the binuclear copper center and allosteric sites on the hemocyanin are affected by heavy metal binding. Cadmium and zinc ions increase the oxygen affinity; mercury and copper ions have the opposite effect. All four heavy metal ions decrease the degree of cooperative oxygen binding. The mercury-induced changes in oxygen binding by 25S Callinectes hemocyanin appear to be the result of that metal's interaction with the high-affinity tryptophan binding site. Mercury binding to the available sulfhydryl group in oxyhemocyanin occurs without functional consequences. Heavy metal, hydrogen, and chloride ions affect the affinity of the first or last oxygen molecules bound to the hemocyanin, which results in the appearance of multiple T (low oxygen affinity) and R (high oxygen affinity) states. Additionally, these ions shift the equilibrium between the low and high oxygen affinity states. The appearance of additional R states at high pH is accompanied by the cleavage of a tyrosine hydrogen bond.

Full Text

Duke Authors

Cited Authors

  • Brouwer, M; Bonaventura, C; Bonaventura, J

Published Date

  • May 1, 1982

Published In

Volume / Issue

  • 21 / 10

Start / End Page

  • 2529 - 2538

PubMed ID

  • 7093201

Electronic International Standard Serial Number (EISSN)

  • 1520-4995

International Standard Serial Number (ISSN)

  • 0006-2960

Digital Object Identifier (DOI)

  • 10.1021/bi00539a037


  • eng