Characteristics of 92 kDa type IV collagenase/gelatinase produced by granulocytic leukemia cells: structure, expression of cDNA in E. coli and enzymatic properties

doi:10.1016/0167-4838(94)90038-8

Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology

Volume 1204, Issue 1, 11 January 1994, Pages 97-107

Biochimica et Biophy...

https://doi.org/10.1016/0167-4838(94)90038-8 Get rights and content

Abstract

Human neutrophils can be triggered to release the collagenolytic metalloenzymes, interstitial collagenase and 92 kDa type IV collagenase/gelatinase. We have isolated and sequenced a 2.3 kb cDNA from a chronic granulocytic leukemia cDNA library that encodes for human neutrophil type IV collagenase. With the exception of one amino-acid substitution at position 280 (Arg → Gln), the deduced amino-acid sequences of neutrophil gelatinase are identical to the amino-acid sequences of the enzyme isolated from fibrosarcoma cells. Expression of the cDNA in E. coli yielded a 72 kDa protein having a gelatinolytic activity on zymogram gel. The recombinant enzyme was activated with APMA and trypsin. The activation was accompanied by a reduction in molecular weight of ≈ 10 kDa; such a reduction is characteristic of matrix metalloproteinases. The recombinant gelatinase cleaved native type V and XI collagens. Native type I collagen was not a substrate for the enzyme. These data suggest that native and recombinant 92 kDa type IV collagenase produced in E. coli have similar biochemical properties. The successful expression of the collagenase in a prokaryotic system will greatly facilitate the structure-function characterization of the enzyme and allow a more precise analysis of its physiological and pathological roles.

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Thus, the results of the meta-analysis published by Wen D et al. must be interpreted with caution. MMP-9 (gelatinase B), a 92 kDa protein is mainly known for cleaving types IV and V collagen protein (Buraczynska et al., 2015; Fenhalls et al., 1999; Pourmotabbed et al., 1994). MMP-9 is highly expressed in atherosclerotic plagues and plays a key role in blood brain barrier disruption (Barr et al., 2010) leading to hemorrhagic transformation (Jha et al., 2014).
Ischemic stroke (IS) is a complex and devastating vascular disease that has become one of the leading causes of disability and mortality worldwide. Several studies have shown the association between matrix metalloproteinase (MMP) family gene polymorphisms and IS. However, the results have been indecisive.
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Our meta-analysis suggests that MMP-9 (−1562C/T) and MMP-12 (−1082 A/G) gene polymorphisms could be a risk factor for IS while MMP-1 (−1607 1G/2G), MMP-2 (−1306C/T) & (−735C/T) and MMP-3 (−1612 5A/6A) have no association with the risk of causing IS. However, large prospective studies with sufficient power are required to validate our findings.
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Characteristics of 92 kDa type IV collagenase/gelatinase produced by granulocytic leukemia cells: structure, expression of cDNA in E. coli and enzymatic properties

Abstract

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

jtJ. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Trends Genet.

J. Biol. Chem.

Blood

Biochim. Biophys. Acta.

Collegan Rel. Res.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Biochem. J.

Biochem. J.