Linked Life Data

2176211

Source:http://linkedlifedata.com/resource/pubmed/id/2176211

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
36
pubmed:dateCreated
1991-2-14
pubmed:abstractText
A chimeric gene composed of the mouse metallothionein promoter linked to the 5' end of the 9.1-kilobase pair rabbit procollagenase (matrix metalloproteinase-1) gene was stably transfected into baby hamster kidney (BHK) cells. Like the native protein, the recombinant procollagenase synthesized and secreted by these cells was the product of a 2.1-kilobase pair transcript which was translated into a procollagenase protein of 57 kDa, with a small amount of protein that co-migrated with the glycosylated form of the native protein at 61 kDa. The BHK cells expressed levels of recombinant procollagenase equal to or exceeding those of rabbit synovial fibroblasts stimulated with phorbol myristate acetate, where procollagenase mRNA may comprise 2% of the mRNA population. Although minimal (approximately 10%) collagenolysis was seen when the zymogen was activated with trypsin or an organ-omercurial compound, the expression of full collagenolytic activity of the recombinant protein depended on the presence of stromelysin (matrix metalloproteinase-3). Purified recombinant collagenase displayed a specific activity of 8,400 units/mg of enzyme (1 unit degraded 1 microgram of collagen/minute at 37 degrees C) when fully activated, which was accomplished by the specific cleavage of the Gln80-Phe81 bond of procollagenase by stromelysin. We conclude that 1) these stably transfected BHK cells represent a high yield source of recombinant mammalian procollagenase, 2) activation of procollagenase depends on the presence of stromelysin, and 3) recombinant procollagenase from this high yield source may be useful in future studies to elucidate the detailed mechanism(s) involved in the activation of this enzyme.
pubmed:grant
pubmed:commentsCorrections
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
265
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
22262-9
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:2176211-Animals, pubmed-meshheading:2176211-Base Sequence, pubmed-meshheading:2176211-Cell Line, pubmed-meshheading:2176211-Chimera, pubmed-meshheading:2176211-Collagenases, pubmed-meshheading:2176211-Enzyme Activation, pubmed-meshheading:2176211-Enzyme Precursors, pubmed-meshheading:2176211-Genes, pubmed-meshheading:2176211-Genetic Vectors, pubmed-meshheading:2176211-Kinetics, pubmed-meshheading:2176211-Matrix Metalloproteinase 3, pubmed-meshheading:2176211-Metalloendopeptidases, pubmed-meshheading:2176211-Metallothionein, pubmed-meshheading:2176211-Microbial Collagenase, pubmed-meshheading:2176211-Molecular Sequence Data, pubmed-meshheading:2176211-Oligonucleotide Probes, pubmed-meshheading:2176211-Plasmids, pubmed-meshheading:2176211-Promoter Regions, Genetic, pubmed-meshheading:2176211-Rabbits, pubmed-meshheading:2176211-Recombinant Proteins, pubmed-meshheading:2176211-Transfection
pubmed:year
1990
pubmed:articleTitle
Rabbit procollagenase synthesized and secreted by a high-yield mammalian expression vector requires stromelysin (matrix metalloproteinase-3) for maximal activation.
pubmed:affiliation
Department of Medicine, Dartmouth Medical School, Hanover, New Hampshire 03756.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't