The mechanisms of inactivation of sulfite oxidase by periodate and arsenite.

Published

Journal Article

The inactivation of sulfite oxidase, a molybdoenzyme containing the Mo cofactor, by arsenite and periodate was investigated. In contrast to ferricyanide (Gardlik, S., and Rajagopalan, K.V. (1991) J. Biol. Chem. 266, 4889-4895), neither of these reagents causes oxidation of the pterin ring of the Mo cofactor. Instead, inactivation by these reagents appears to involve attack on sulfhydryl groups at the active site of the enzyme. The inactivation of sulfite oxidase by arsenite was shown to be dependent on the presence of O2 and on the enzymatic oxidation of arsenite to arsenate. The inactivation was preventable by the presence of sulfite, or by the use of cytochrome c as the electron acceptor instead of O2. It is concluded that inactivation by arsenite is the result of arsenite displacement of Mo during enzymatic oxidation of arsenite to arsenate, when Mo transiently breaks its bond to protein or molybdopterin sulfhydryl(s) in order to provide a site for transfer of electrons to O2. Data indicate that arsenite is properly oriented to displace Mo only once every 20,800 turnovers, thus accounting for the slow rate of inactivation by this reagent. Inactivation of sulfite oxidase by periodate is believed to occur as the result of direct attack of periodate on the thiolate ligands of Mo, either those of the protein and/or molybdopterin, leading to Mo loss. Treatment of enzyme with even low levels of periodate resulted in loss of Mo and both sulfite:cytochrome c and sulfite:O2 activities. Molybdopterin of periodate-inactivated enzyme retained the ability to reconstitute nitrate reductase apoprotein in nit-1 extracts and the ability to reduce dichlorophenolindophenol, indicating that the pterin ring had not been oxidized.

Full Text

Duke Authors

Cited Authors

  • Gardlik, S; Rajagopalan, KV

Published Date

  • September 5, 1991

Published In

Volume / Issue

  • 266 / 25

Start / End Page

  • 16627 - 16632

PubMed ID

  • 1653244

Pubmed Central ID

  • 1653244

International Standard Serial Number (ISSN)

  • 0021-9258

Language

  • eng

Conference Location

  • United States