Linked Life Data

7663162

Source:http://linkedlifedata.com/resource/pubmed/id/7663162

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1995-10-12
pubmed:abstractText
A rat cDNA encoding phospholipase C-gamma 1 (PLC-gamma 1) was expressed as a histidine-tagged fusion protein in insect cells utilizing the expression vector pBlueBacHis. The fusion protein was purified by Ni(2+)-agarose affinity chromatography to apparent homogeneity as defined by SDS-PAGE and Coomassie staining. Using a Triton X-100/PIP2 mixed micelle assay, (His6)-PLC-gamma 1 exhibited a calcium-dependent specific enzyme activity of 3 mumol/min/mg, a value similar to that reported for purified bovine brain PLC-gamma 1. Also similar to bovine brain PLC-gamma 1, (His6)-PLC-gamma 1 activity was stimulated by phosphatidic acid and inhibited by adenosine 5'monophosphate. (His6)-PLC-gamma 1 interacted directly with two known PLC-gamma 1 binding proteins, dynamin and the activated EGF receptor. Also, purified (His6)-PLC-gamma 1 was a tyrosine phosphorylation substrate for purified EGF receptor. These results suggest that (His6)-PLC-gamma 1 can be overexpressed as a functional enzyme in baculovirus-infected insect cells and purified by a one-step metal affinity chromatography procedure.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
1046-5928
pubmed:author
pubmed:issnType
Print
pubmed:volume
6
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
278-83
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed:year
1995
pubmed:articleTitle
Baculovirus expression and purification of the second messenger enzyme phospholipase C-gamma 1, a tyrosine kinase substrate.
pubmed:affiliation
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.