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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1990-4-26
|
| pubmed:abstractText |
Although the optimum substrate for lipoamidase (lipoyl-X hydrolase) has not yet been determined, it is known that lipoamidase activity, as determined by hydrolysis of the synthetic substrate lipoyl 4-aminobenzoate (LPAB), is widely distributed in pig brain tissues, i.e. in the cerebrum, cerebellum and medulla. Over 95% of the enzyme activity is present in the membrane subfractions, indicating that brain lipoamidase is an integral membrane protein enzyme. To elucidate the chemical nature and the optimum substrate of the abundant lipoamidase in the brain, we isolated it from the membrane subfractions. After an 8-step purification procedure, brain lipoamidase was purified 601-fold and identified as a 140 kDa glycoprotein by SDS/PAGE. A mechanistic study to determine Km and Vmax, values was carried out using various synthetic compounds. Lipoyl-lysine, which is generally believed to be a naturally occurring substrate of lipoamidase, was first compared with biotinyl-lysine, because these two vitamins have reactive sulphur atoms and are similar in molecular mass and structure. The Km for lipoyl-lysine was 333 microM, whereas biotinyl-lysine was not hydrolysed. Stringent specificity for the lipoyl moiety is demonstrated, as expected. Dipeptides of amino acid-lysine structures were studied, and dipeptides of aspartyl- and glutamyl-lysine hydrolysis occurred at high Km (3 mM) values. Thus lysine in the moiety is not very effective as an optimum substrate. The chemical bond structures of the amide bond (lipoyl-lysine) and peptide bond (aspartyl-lysine) were hydrolysed. Next, the ester bond compound was tested, and it was observed that lipolylmethyl ester was hydrolysed at high specificity. These findings indicate that this enzyme has broad specificities with respect to bond structure; it therefore is a unique hydrolase having stringent specificity for lipoic acid and relatively broad specificity for the chemical bond and the X moiety. Various inhibitors were tested; a few reagents, such as organic mercurials, di-isopropylfluorophosphate, 1,10-phenanthroline, sodium azide and angiotensin-converting enzyme inhibitor exhibited some inhibition (not more than 60%). Thus the active centre of this enzyme is a complex type. Although ATP is not hydrolysed and the lowest Km value is exhibited by the synthetic substrate reduced from LPAB (12 microM), some other compounds may still be expected to be hydrolysed by this unique and abundant brain lipoamidase.
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| pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-130084,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-13549405,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-13610913,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-13862582,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-14086741,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-14340769,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-14907713,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-180454,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-2503531,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-2730918,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-3024571,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-3168970,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-3196715,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-3443663,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-416517,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-4502942,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-4581500,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-4862967,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-6180829,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-6418873,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-6769358,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2317196-6783978
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0264-6021
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
1
|
| pubmed:volume |
266
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
427-34
|
| pubmed:dateRevised |
2009-11-18
|
| pubmed:meshHeading |
pubmed-meshheading:2317196-Amidohydrolases,
pubmed-meshheading:2317196-Amino Acids,
pubmed-meshheading:2317196-Animals,
pubmed-meshheading:2317196-Brain,
pubmed-meshheading:2317196-Glycoproteins,
pubmed-meshheading:2317196-Hydrogen-Ion Concentration,
pubmed-meshheading:2317196-Isoelectric Point,
pubmed-meshheading:2317196-Kinetics,
pubmed-meshheading:2317196-Molecular Weight,
pubmed-meshheading:2317196-Subcellular Fractions,
pubmed-meshheading:2317196-Substrate Specificity,
pubmed-meshheading:2317196-Swine,
pubmed-meshheading:2317196-Tissue Distribution
|
| pubmed:year |
1990
|
| pubmed:articleTitle |
Lipoamidase (lipoyl-X hydrolase) from pig brain.
|
| pubmed:affiliation |
Division of Metabolism, National Children's Medical Research Center, Tokyo, Japan.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|