| pubmed-article:8280053 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:8280053 | lifeskim:mentions | umls-concept:C0020331 | lld:lifeskim |
| pubmed-article:8280053 | lifeskim:mentions | umls-concept:C1280500 | lld:lifeskim |
| pubmed-article:8280053 | lifeskim:mentions | umls-concept:C1516893 | lld:lifeskim |
| pubmed-article:8280053 | pubmed:dateCreated | 1994-2-7 | lld:pubmed |
| pubmed-article:8280053 | pubmed:abstractText | (R)-3-Hydroxybutyrate dehydrogenase (BDH) is a phosphatidylcholine-requiring tetrameric enzyme with two thiol groups (SH-1 and SH-2) per protomer. By first protecting the more rapidly reacting thiol group (SH-1) with diamide [1,1'-azobis-(NN'-dimethylformamide), DM] to form DM(SH-1)BDH, SH-2 can be selectively derivatized by reaction with maleimide reagents such as 4-maleimido-2,2,6,6-tetramethyl-piperidine-N-oxyl (MSL), which gives DM(SH-1)MSL(SH-2)BDH. Reduction with dithiothreitol (DTT) regenerates SH-1, yielding MAL(SH-2)BDH (where MAL is the diamagnetic reduction product of MSL-BDH and DTT). The enzymic activity of DM(SH-1)BDH is decreased to approx. 4% relative to the native purified enzyme, and the apparent Km for substrate, KmBOH, is increased approx. 100-fold. Reduction of DM(SH-1)BDH with DTT regenerates SH-1 and restores normal enzymic function. Modification of SH-2 with piperidinylmaleimide [MAL(SH-2)BDH] diminishes enzymic activity to approx. 35% of its original value, but has no significant effect on apparent KmBOH. The doubly derivatized enzyme, DM(SH-1)MSL(SH-2)BDH, has lower enzymic activity [about half that for DM(SH-2)BDH] and a yet higher apparent KmBOH than DM(SH-1)BDH. Derivatization of SH-2 with different maleimide reagents results in diminished activity approximately proportional to the size of the maleimide substituent, suggesting that this inhibition is steric. Whereas modification of SH-1 results in marked changes in kinetic parameters (increased apparent Km and reduced apparent Vmax), derivatization of SH-2 has a lesser effect on enzymic function. Thus SH-1 is postulated to be closer to the active centre than is SH-2, although neither is involved in catalysis, since: (1) the activity of the derivatized enzyme is not abolished; and (2) activity can be enhanced by increasing substrate (and cofactor) concentrations. | lld:pubmed |
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| pubmed-article:8280053 | pubmed:language | eng | lld:pubmed |
| pubmed-article:8280053 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:8280053 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:8280053 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:8280053 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
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| pubmed-article:8280053 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:8280053 | pubmed:month | Dec | lld:pubmed |
| pubmed-article:8280053 | pubmed:issn | 0264-6021 | lld:pubmed |
| pubmed-article:8280053 | pubmed:author | pubmed-author:FleischerSS | lld:pubmed |
| pubmed-article:8280053 | pubmed:author | pubmed-author:McIntyreJ OJO | lld:pubmed |
| pubmed-article:8280053 | pubmed:author | pubmed-author:DaltonL ALA | lld:pubmed |
| pubmed-article:8280053 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:8280053 | pubmed:day | 15 | lld:pubmed |
| pubmed-article:8280053 | pubmed:volume | 296 ( Pt 3) | lld:pubmed |
| pubmed-article:8280053 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:8280053 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:8280053 | pubmed:pagination | 563-9 | lld:pubmed |
| pubmed-article:8280053 | pubmed:dateRevised | 2009-11-18 | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:meshHeading | pubmed-meshheading:8280053-... | lld:pubmed |
| pubmed-article:8280053 | pubmed:year | 1993 | lld:pubmed |
| pubmed-article:8280053 | pubmed:articleTitle | Effect of selective thiol-group derivatization on enzyme kinetics of (R)-3-hydroxybutyrate dehydrogenase. | lld:pubmed |
| pubmed-article:8280053 | pubmed:affiliation | Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235. | lld:pubmed |
| pubmed-article:8280053 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:8280053 | pubmed:publicationType | Research Support, U.S. Gov't, P.H.S. | lld:pubmed |
| pubmed-article:8280053 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
