Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
|
pubmed:dateCreated |
1989-5-10
|
pubmed:abstractText |
A modified procedure was used to purify sulfite oxidase (sulfite:O2 oxidoreductase; EC 1.8.3.1) from chicken liver in high yield. The modifications included dialysis of the enzyme against a buffered solution containing sodium molybdate (prior to ion-exchange chromatography), which apparently reconstituted any demolybdo enzyme present in the extract, and phenyl-Sepharose column chromatography. Analysis showed that the purified enzyme contained Mo and heme in a 1.03:1.00 ratio, indicating that the enzyme was prosthetically intact; exogenous heme and other colored proteins were absent from the final pool. Treatment of the sulfite-reduced enzyme with 50 mM cyanide at pH 8.5 resulted in a gradual loss of catalytic activity with a half-life of 19.7 min. Analysis of the cyanide-inactivated enzyme gave a Mo:heme ratio of 1.02:1.00, providing the first direct evidence that the enzyme does not lose molybdenum when inactivated with cyanide. This modified purification procedure provides enzyme in high yield which is well-suited for experiments requiring prosthetically intact enzyme and which is not contaminated with extraneous heme or with other redox active proteins.
|
pubmed:grant | |
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Apr
|
pubmed:issn |
0003-9861
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:volume |
270
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
383-90
|
pubmed:dateRevised |
2009-11-19
|
pubmed:meshHeading |
pubmed-meshheading:2930196-Animals,
pubmed-meshheading:2930196-Buffers,
pubmed-meshheading:2930196-Chemical Precipitation,
pubmed-meshheading:2930196-Chickens,
pubmed-meshheading:2930196-Chromatography, Gel,
pubmed-meshheading:2930196-Chromatography, Ion Exchange,
pubmed-meshheading:2930196-Cyanides,
pubmed-meshheading:2930196-Dialysis,
pubmed-meshheading:2930196-Hot Temperature,
pubmed-meshheading:2930196-Liver,
pubmed-meshheading:2930196-Oxidoreductases,
pubmed-meshheading:2930196-Oxidoreductases Acting on Sulfur Group Donors,
pubmed-meshheading:2930196-Quality Control
|
pubmed:year |
1989
|
pubmed:articleTitle |
Purification of prosthetically intact sulfite oxidase from chicken liver using a modified procedure.
|
pubmed:affiliation |
Department of Chemistry, University of Arizona, Tucson 85721.
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|