Linked Life Data

1651335

Source:http://linkedlifedata.com/resource/pubmed/id/1651335

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
23
pubmed:dateCreated
1991-9-13
pubmed:abstractText
Phosphoinositide-specific phospholipase C (PLC) activities have been partially purified from cultured vascular smooth muscle cells and analyzed for substrate specificity, calcium and pH requirements, and molecular weight. The purification procedure involved DEAE-cellulose and heparin-Sepharose chromatographies followed by Mono Q and size exclusion high performance liquid chromatography. This technique resolves multiple peaks of activity using phosphatidylinositol (PI) and PI 4,5-bisphosphate (PIP2) as substrates. The major peak was purified to near homogeneity as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PLC activity in vascular smooth muscle cells can be divided into two types based on their calcium and pH requirements, substrate preferences, and molecular weights. The low molecular weight PLC hydrolyzes both PI and PIP2, has a molecular mass of 58 kDa, requires the most calcium for full activation, and has a PI-pH profile that shifts slightly with calcium concentration. Screening a cDNA library with oligonucleotides directed against several of the known PLCs identified a highly expressed PLC cDNA that is 99% homologous to PLC-alpha, suggesting that this low molecular weight peak in fact corresponds to PLC-alpha. The high molecular mass peak (157 kDa) shows much greater activity against PI than PIP2, is active at lower calcium concentrations, and has a PI-pH optimum of 5.0 regardless of calcium concentration. Each of the PIP2 PLC activities is strongly dependent on the relative levels of calcium and pH in the assay buffer. These observations suggest that vascular smooth muscle contains both a high and low molecular weight PLC whose activities are affected markedly by the changes in calcium and pH accompanying hormonal stimulation of the cell.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
266
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
15498-504
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:1651335-Animals, pubmed-meshheading:1651335-Blotting, Northern, pubmed-meshheading:1651335-Calcium, pubmed-meshheading:1651335-Cells, Cultured, pubmed-meshheading:1651335-Chromatography, Liquid, pubmed-meshheading:1651335-Cloning, Molecular, pubmed-meshheading:1651335-DNA, pubmed-meshheading:1651335-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:1651335-Hydrogen-Ion Concentration, pubmed-meshheading:1651335-Hydrolysis, pubmed-meshheading:1651335-Isoenzymes, pubmed-meshheading:1651335-Molecular Weight, pubmed-meshheading:1651335-Muscle, Smooth, Vascular, pubmed-meshheading:1651335-Phosphatidylinositol Diacylglycerol-Lyase, pubmed-meshheading:1651335-Phosphoinositide Phospholipase C, pubmed-meshheading:1651335-Phosphoric Diester Hydrolases, pubmed-meshheading:1651335-RNA, Messenger, pubmed-meshheading:1651335-Rats
pubmed:year
1991
pubmed:articleTitle
Characterization of phosphatidylinositol-specific phospholipase C from cultured vascular smooth muscle cells.
pubmed:affiliation
Department of Medicine, Emory University, Atlanta, Georgia 30322.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't