Purification of Dihydroxyacetone Phosphate Acyltransferase from Guinea Pig Liver Peroxisomes
dc.contributor.author | Webber K. O. , | en_US |
dc.contributor.author | Hajra A. K. , | en_US |
dc.date.accessioned | 2006-04-10T15:56:38Z | |
dc.date.available | 2006-04-10T15:56:38Z | |
dc.date.issued | 1993-01 | en_US |
dc.identifier.citation | Webber K. O., , Hajra A. K., (1993/01)."Purification of Dihydroxyacetone Phosphate Acyltransferase from Guinea Pig Liver Peroxisomes." Archives of Biochemistry and Biophysics 300(1): 88-97. <http://hdl.handle.net/2027.42/31045> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6WB5-45PTR3C-FR/2/443523232d969343ae67d29ed31f44a7 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/31045 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=8380974&dopt=citation | en_US |
dc.description.abstract | Dihydroxyacetone phosphate acyltransferase (EC 2.3.1.42), a peroxisomal enzyme which initiates the biosynthesis of glycerolipids (especially the ether-linked glycerolipids) in higher eukaryotes, has been purified by over 3250-fold from guinea pig liver. Initial stages of purification entailed isolation of liver peroxisomes by a combination of differential and density-gradient centrifugation. Dihydroxyacetone phosphate acyltransferase was solubilized from peroxisomal membranes with 3-[3-cholamidopropyl)dimethylammonio]-1-propane sulfonate at moderate ionic strength (0.15 M NaCl). The solubilized enzyme was further purified by a regimen of size-exclusion chromatography, cation-exchange chromatography, and hydroxylapatite chromatography. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of different fractions during the purification of the enzyme, a 69-kDa protein band copurified with the enzyme activity, indicating that the monomeric enzyme may have a Mr of 69,000. This was verified by further purifying the enzyme by chromatofocusing, when a single 69-kDa band was observed on SDS-PAGE. The Mr of dihydroxyacetone phosphate acyltransferase determined by gel filtration is 90 kDa. The Vmax of the purified enzyme was 4 pmol acyldihydroxyacetone phosphate (acylDHAP) formed per minute per milligram protein and the Km(DHAP) is 70 [mu]M when assayed at saturating concentrations of palmitoylCoA. Free coenzyme A inhibits the acyltransferase reaction with an inhibition constant (Ki) of approximately 0.76 mM. To date, this is the most highly purified DHAP acyltransferase(>3200-fold) of mammalian origin. | en_US |
dc.format.extent | 978120 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | Purification of Dihydroxyacetone Phosphate Acyltransferase from Guinea Pig Liver Peroxisomes | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Public Health | en_US |
dc.subject.hlbsecondlevel | Chemistry | en_US |
dc.subject.hlbsecondlevel | Chemical Engineering | en_US |
dc.subject.hlbsecondlevel | Biological Chemistry | en_US |
dc.subject.hlbtoplevel | Engineering | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Biological Chemistry, University of Michigan, Mental Health Research Institute, Neuroscience Lab, Ann Arbor, MI 48109, USA | en_US |
dc.contributor.affiliationum | Department of Biological Chemistry, University of Michigan, Mental Health Research Institute, Neuroscience Lab, Ann Arbor, MI 48109, USA | en_US |
dc.identifier.pmid | 8380974 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/31045/1/0000722.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1006/abbi.1993.1013 | en_US |
dc.identifier.source | Archives of Biochemistry and Biophysics | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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