Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
10
|
pubmed:dateCreated |
1992-4-28
|
pubmed:abstractText |
The human 72-kDa (CLG4A) and 92-kDa (CLG4B) type IV collagenases contain a domain consisting of three contiguous copies of the fibronectin (FN)-derived type II homology unit (T2HU), T2HU-1, T2HU-2, and T2HU-3. To investigate the functional role of this domain, we have constructed plasmids expressing beta-galactosidase fusion proteins with one or more of the CLG4B-derived T2HU. The gelatin binding assays demonstrate that a single copy of T2HU-2 renders beta-galactosidase capable of binding gelatin. The three repeats, however, differ dramatically in their capacity to bind gelatin, with T2HU-1 and T2HU-3 having significantly less binding activity than T2HU-2. Using alanine scanning mutagenesis we have defined the amino acid residues (Arg307, Asp309, Asn319, Tyr320, Asp323) that are critical for gelatin binding of T2HU-2. The low gelatin binding of T2HU-1 compared to T2HU-2 was traced to the non-conserved residues Ala228-Ala and Leu253-Pro. The results suggest that the gelatin binding of the type IV collagenase proenzyme is mediated by the FN-like domain, although the presence of another gelatin-binding site cannot be excluded. The FN domain-mediated binding, however, is not a rate-limiting step in the hydrolysis of gelatin by the enzyme.
|
pubmed:grant | |
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Alanine,
http://linkedlifedata.com/resource/pubmed/chemical/Dimethyl Sulfoxide,
http://linkedlifedata.com/resource/pubmed/chemical/Fibronectins,
http://linkedlifedata.com/resource/pubmed/chemical/Gelatin,
http://linkedlifedata.com/resource/pubmed/chemical/Microbial Collagenase,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/beta-Galactosidase
|
pubmed:status |
MEDLINE
|
pubmed:month |
Apr
|
pubmed:issn |
0021-9258
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:day |
5
|
pubmed:volume |
267
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
6776-81
|
pubmed:dateRevised |
2007-11-14
|
pubmed:meshHeading |
pubmed-meshheading:1313021-Alanine,
pubmed-meshheading:1313021-Amino Acid Sequence,
pubmed-meshheading:1313021-Binding Sites,
pubmed-meshheading:1313021-Dimethyl Sulfoxide,
pubmed-meshheading:1313021-Escherichia coli,
pubmed-meshheading:1313021-Fibronectins,
pubmed-meshheading:1313021-Gelatin,
pubmed-meshheading:1313021-Humans,
pubmed-meshheading:1313021-Microbial Collagenase,
pubmed-meshheading:1313021-Molecular Sequence Data,
pubmed-meshheading:1313021-Mutagenesis,
pubmed-meshheading:1313021-Plasmids,
pubmed-meshheading:1313021-Recombinant Fusion Proteins,
pubmed-meshheading:1313021-Repetitive Sequences, Nucleic Acid,
pubmed-meshheading:1313021-Sequence Alignment,
pubmed-meshheading:1313021-beta-Galactosidase
|
pubmed:year |
1992
|
pubmed:articleTitle |
Alanine scanning mutagenesis and functional analysis of the fibronectin-like collagen-binding domain from human 92-kDa type IV collagenase.
|
pubmed:affiliation |
Division of Dermatology, Washington University School of Medicine, St. Louis, Missouri 63110.
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
|