| dc.contributor.author |
Labrou, NE |
en |
| dc.date.accessioned |
2014-06-06T06:44:16Z |
|
| dc.date.available |
2014-06-06T06:44:16Z |
|
| dc.date.issued |
2000 |
en |
| dc.identifier.issn |
87555093 |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/1775 |
|
| dc.relation.uri |
http://www.scopus.com/inward/record.url?eid=2-s2.0-0033882973&partnerID=40&md5=052464e4058ae73f7214a344774894ad |
en |
| dc.subject |
Affinity labelling |
en |
| dc.subject |
Alcohol dehydrogenase |
en |
| dc.subject |
Anthraquinone dyes |
en |
| dc.subject |
Triazine dyes |
en |
| dc.subject.other |
alcohol dehydrogenase |
en |
| dc.subject.other |
dye |
en |
| dc.subject.other |
triazine derivative |
en |
| dc.subject.other |
unclassified drug |
en |
| dc.subject.other |
vilmafix blue a r |
en |
| dc.subject.other |
article |
en |
| dc.subject.other |
binding affinity |
en |
| dc.subject.other |
binding site |
en |
| dc.subject.other |
concentration (parameters) |
en |
| dc.subject.other |
controlled study |
en |
| dc.subject.other |
enzyme binding |
en |
| dc.subject.other |
enzyme inactivation |
en |
| dc.subject.other |
gel filtration chromatography |
en |
| dc.subject.other |
nonhuman |
en |
| dc.subject.other |
pH |
en |
| dc.subject.other |
priority journal |
en |
| dc.subject.other |
purification |
en |
| dc.subject.other |
reversed phase high performance liquid chromatography |
en |
| dc.subject.other |
temperature |
en |
| dc.subject.other |
thin layer chromatography |
en |
| dc.subject.other |
yeast |
en |
| dc.subject.other |
Affinity Labels |
en |
| dc.subject.other |
Alcohol Dehydrogenase |
en |
| dc.subject.other |
Anthraquinones |
en |
| dc.subject.other |
Chromatography, High Pressure Liquid |
en |
| dc.subject.other |
Chromatography, Thin Layer |
en |
| dc.subject.other |
Coloring Agents |
en |
| dc.subject.other |
Models, Molecular |
en |
| dc.subject.other |
Molecular Structure |
en |
| dc.subject.other |
NAD |
en |
| dc.subject.other |
Protein Binding |
en |
| dc.subject.other |
Protein Conformation |
en |
| dc.subject.other |
Saccharomyces |
en |
| dc.subject.other |
Structure-Activity Relationship |
en |
| dc.subject.other |
Triazines |
en |
| dc.title |
Dye affinity labelling of yeast alcohol dehydrogenase |
en |
| heal.type |
journalArticle |
en |
| heal.publicationDate |
2000 |
en |
| heal.abstract |
The interaction of yeast alcohol dehydrogenase (ADH) with the reactive chlorotriazine dye Vilmafix Blue A-R (VBAR) was studied. VBAR was purified to homogeneity on lipophilic Sephadex LH-20 and characterised by reverse phase HPLC and analytical TLC. Incubation of ADH with purified VBAR at pH 8.0 and 37°C resulted in a time-dependent inactivation of the enzyme. The observed rate of enzyme inactivation (k(obs)) exhibited a non-linear dependence on VBAR concentration from 22 to 106 nmol, with a maximum rate of inactivation (k 3) of 0.134min -1 and k(D) of 141.7 μM. The inhibition was irreversible and activity could not be recovered by gel-filtration chromatography. The inactivation of ADH by VBAR was competitively inhibited by the nucleotides NADH and NAD +. These results suggest that VBAR acts as an affinity label at the nucleotide binding site of yeast ADH. |
en |
| heal.journalName |
Journal of Enzyme Inhibition |
en |
| dc.identifier.issue |
5 |
en |
| dc.identifier.volume |
15 |
en |
| dc.identifier.spage |
487 |
en |
| dc.identifier.epage |
496 |
en |