Linked Life Data

8363589

Source:http://linkedlifedata.com/resource/pubmed/id/8363589

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
1993-9-24
pubmed:abstractText
cDNAs corresponding to the precursor and mature forms of rat carnitine palmitoyltransferase II (CPT II) were found to be readily expressed in Escherichia coli. In both cases, catalytically active immunoreactive protein was produced and became largely membrane-associated. The precursor form of the enzyme was not proteolytically processed. Removal of 126 bp from the 5' end of the cDNA coding region allowed expression of a truncated CPT II (lacking the N-terminal 17 residues of the mature protein), but this product was inactive. cDNAs encoding the precursor and mature forms of human CPT II resisted direct expression in E. coli. However, the impediment was overcome when the latter cDNA was ligated in-frame 3' to sequence encoding a glutathione S-transferase. This construct yielded abundant quantities of the corresponding fusion protein, a portion of which was soluble and catalytically active. In vitro transcription and translation of the various cDNAs established that the lower mobility on SDS/PAGE of rat CPT II compared with its human counterpart (despite their identical numbers of amino acids) is an intrinsic property of the primary sequences of the proteins themselves. Also, the human cDNA was found to contain an artifactual termination signal for T3 RNA polymerase that could be bypassed by the T7 polymerase. Thus rat CPT II can be expressed in active form in E. coli with characteristics similar to those of the enzyme in mitochondria, opening the way to future location of active sites within the molecule. An alternative expression system will be needed for similar studies on human CPT II.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1417736, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1439783, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-14907713, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1600836, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1869564, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1961767, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-1988962, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2016062, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2109307, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2355017, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2355018, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2656156, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-2897629, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-3597441, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-3597442, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-6157353, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-6615466, http://linkedlifedata.com/resource/pubmed/commentcorrection/8363589-8449948
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0264-6021
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
294 ( Pt 1)
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
79-86
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Expression of precursor and mature carnitine palmitoyltransferase II in Escherichia coli and in vitro: differential behaviour of rat and human isoforms.
pubmed:affiliation
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas 75235.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.