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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
28
|
| pubmed:dateCreated |
1993-11-24
|
| pubmed:databankReference | |
| pubmed:abstractText |
We have characterized inositol phospholipid-specific phospholipase C (PLC) isozymes in bovine retina. Chromatography of a retinal homogenate on a heparin column partially resolved six peaks of PLC activity, which differed in their relative selectivities for the substrates phosphatidyl 4,5-bisphosphate (PIP2) and phosphatidylinositol (PI). Five of the peaks were shown to correspond to the known PLC isozymes PLC-beta 1, PLC-beta 3, PLC-gamma 1, PLC-delta 1, and PLC-delta 2. PLC-beta 1, PLC-beta 3, PLC-gamma 1, and PLC-delta 1 in the retinal fractions were identified by immunoblotting with isozyme-specific antibodies, and PLC-delta 2 was identified by direct sequencing of tryptic peptides. PLC-gamma 2 and PLC-beta 2 were not detectable by immunoblot analysis. In addition to five of the seven mammalian PLC isozymes identified to date, bovine retina contained a previously unidentified PLC, which exhibited the highest selectivity for PIP2 over PI. The new PLC was purified from a retinal particulate fraction to yield a preparation that contained a major protein band with an apparent molecular mass of 130 kDa on SDS-polyacrylamide gels. Sequence analysis of 12 tryptic peptides derived from the 130-kDa protein suggested that the primary structure of the new PLC is similar to those members of beta-type PLC isozymes, especially to that of PLC-norpA, which was originally identified in Drosophila eye. The new enzyme was thus named PLC-beta 4. A search of a rat brain cDNA library with the polymerase chain reaction and oligonucleotide primers based on common PLC amino acid sequences resulted in the cloning of a rat brain cDNA corresponding to a previously uncharacterized PLC. The cDNA encoded a putative polypeptide of 1176 amino acids, with a calculated molecular mass of 134,532 daltons, that contained the sequences of all 12 tryptic peptides of PLC-beta 4. Furthermore, the deduced amino acid sequence of the encoded protein was more related to PLC-norpA than to any of the three mammalian PLC-beta isozymes. These results suggest that the brain cDNA encodes PLC-beta 4, which is likely a mammalian homolog of PLC-norpA.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Complementary,
http://linkedlifedata.com/resource/pubmed/chemical/Isoenzymes,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphatidylinositol 4,5-Diphosphate,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphatidylinositol Phosphates,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphatidylinositols,
http://linkedlifedata.com/resource/pubmed/chemical/Type C Phospholipases
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Oct
|
| pubmed:issn |
0021-9258
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
5
|
| pubmed:volume |
268
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
21318-27
|
| pubmed:dateRevised |
2007-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:8407970-Amino Acid Sequence,
pubmed-meshheading:8407970-Animals,
pubmed-meshheading:8407970-Base Sequence,
pubmed-meshheading:8407970-Blotting, Western,
pubmed-meshheading:8407970-Cattle,
pubmed-meshheading:8407970-Chromatography, Gel,
pubmed-meshheading:8407970-Chromatography, High Pressure Liquid,
pubmed-meshheading:8407970-Cloning, Molecular,
pubmed-meshheading:8407970-DNA, Complementary,
pubmed-meshheading:8407970-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8407970-Isoenzymes,
pubmed-meshheading:8407970-Molecular Sequence Data,
pubmed-meshheading:8407970-Peptide Mapping,
pubmed-meshheading:8407970-Phosphatidylinositol 4,5-Diphosphate,
pubmed-meshheading:8407970-Phosphatidylinositol Phosphates,
pubmed-meshheading:8407970-Phosphatidylinositols,
pubmed-meshheading:8407970-Polymerase Chain Reaction,
pubmed-meshheading:8407970-Retina,
pubmed-meshheading:8407970-Sequence Homology, Amino Acid,
pubmed-meshheading:8407970-Substrate Specificity,
pubmed-meshheading:8407970-Type C Phospholipases
|
| pubmed:year |
1993
|
| pubmed:articleTitle |
Purification, molecular cloning, and sequencing of phospholipase C-beta 4.
|
| pubmed:affiliation |
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|