| Stimulation of polyprenyl 4-hydroxybenzoate transferase activity by sodium cholate and 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate. | |
| | |
MedLine Citation:
|
PMID: 16643834 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Polyprenyl 4-hydroxybenzoate transferase (Coq2p) plays a central role in ubiquinone biosynthesis. Coq2p mediates the conjugation of 4-hydroxybenzoate, the benzoquinone ring precursor, with the completed side chain. The activity is most easily assayed by measuring the rate of incorporation of 4-hydroxybenzoate as radiolabeled substrate into polyprenyl 4-hydroxybenzoate. The in vitro assay requires addition of a detergent into the reaction mixture to activate enzyme activity, and Triton X-100 is used for this purpose in the routine assay. We have found that both 3-[(cholamidopropyl)dimethylammonio]-1-propanesulfonate and sodium cholate, but not sodium deoxycholate, lysophosphatidyl choline, or octylglucoside, significantly stimulate the activity over that measured with Triton X-100. High-performance liquid chromatography analysis of lipid extracts revealed that the increase of specific activity resulted in a similar increase in reaction product, this effect is due not merely to a better lipid extraction but also to the actual stimulation of enzyme activity. With our improved method, we were able to measure Coq2p activity with much greater sensitivity in both fresh and frozen/thawed mitochondria and in crude homogenates obtained from cultured cells. Our method will simplify evaluation of Coq2p activity in scarce biological materials, such as cells obtained from human tissue biopsies, and thus it will facilitate the biochemical characterization of ubiquinone deficiencies. |
| | |
Authors:
|
María I Burón; María D Hermán; Francisco J Alcaín; José M Villalba |
Related Documents
:
|
1163994 - Proline biosynthesis and degradation in mammalian cells and tissue. 3062294 - A sensitive radioenzymatic assay for epinephrine forming enzymes. 3896024 - Isocitrate dehydrogenase assays on intact bacterial cells. 10699674 - The measurement of toluene dioxygenase activity in biofilm culture of pseudomonas putid... 15258194 - Niacin noncompetitively inhibits dgat2 but not dgat1 activity in hepg2 cells. 10098864 - Metabolic consequences of the cytochrome c oxidase deficiency in brain of copper-defici... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2006-04-18 |
Journal Detail:
|
Title: Analytical biochemistry Volume: 353 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 2006 Jun |
Date Detail:
|
Created Date: 2006-05-15 Completed Date: 2006-08-01 Revised Date: 2006-11-15 |
Medline Journal Info:
|
Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: United States |
Other Details:
|
Languages: eng Pagination: 15-21 Citation Subset: IM |
Affiliation:
|
Departamento de Biología Celular, Fisiología e Inmunología, Facultad de Ciencias, Campus Universitario de Rabanales, Edificio Severo Ochoa, Universidad de Córdoba, Spain. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Alkyl and Aryl Transferases
/
drug effects* Animals Cells, Cultured Cholic Acids / pharmacology* Chromatography, High Pressure Liquid Chromatography, Thin Layer Detergents / pharmacology* HL-60 Cells Humans Lipids / chemistry Mitochondria, Liver / enzymology Octoxynol / pharmacology Parabens / metabolism Rats Sodium Cholate / pharmacology* Ubiquinone / biosynthesis* |
| Chemical | |
Reg. No./Substance:
|
0/Cholic Acids; 0/Detergents; 0/Lipids; 0/Parabens; 1339-63-5/Ubiquinone; 361-09-1/Sodium Cholate; 75621-03-3/3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; 9002-93-1/Octoxynol; 99-96-7/4-hydroxybenzoic acid; EC 2.5.-/Alkyl and Aryl Transferases; EC 2.5.1.-/4-hydroxybenzoate polyprenyltransferase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Aluminum determination in biological fluids and dialysis concentrates via chelation with 8-hydroxyqu...
Next Document: Development of a high-throughput purification method and a continuous assay system for chlorophyllas...