Purification of a Type V Collagen Degrading Metalloproteinase from Rabbit Alveolar Macrophages

Carlo L. Mainardi, Margaret S. Hibbs, Karen Hasty, Jerome M. Seyer

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

A proteinase capable of degrading the helical region of native human type V collagen was identified in serum-free culture medium from “in vivo-activated” rabbit alveolar macrophages. This enzyme was purified to homogeneity using a combination of gelfiltration, ion-exchange and affinity chromatography. Analysis of the purified material by gel electrophoresis revealed a single broad band with relative molecular mass of 82,000 daltons Enzyme activity was eluted from the gel in a region corresponding to the stained band. The protein band was also found to stain positively using the periodic acid-Schiff technique indicating that it was glycosylated. Amino acid analysis revealed a composition rich in acidic residues. The enzyme cleaved type V collagen into three large molecular weight doublets consistent with two cleavage sites within the helix but was inactive against native type I collagen. However, when type I or type V collagen was heat-denatured, the enzyme degraded the alpha chains to small molecular weight peptides.

Original languageEnglish (US)
Pages (from-to)479-492
Number of pages14
JournalTopics in Catalysis
Volume4
Issue number6
DOIs
StatePublished - Jan 1 1984

Fingerprint

Collagen Type V
Alveolar Macrophages
Metalloproteases
Collagen
Purification
Rabbits
Enzymes
Gels
Molecular weight
Affinity chromatography
Periodic Acid
Molecular Weight
Serum-Free Culture Media
Enzyme activity
Molecular mass
Collagen Type I
Electrophoresis
Ion exchange
Ion Exchange Chromatography
Peptide Hydrolases

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Rheumatology

Cite this

Purification of a Type V Collagen Degrading Metalloproteinase from Rabbit Alveolar Macrophages. / Mainardi, Carlo L.; Hibbs, Margaret S.; Hasty, Karen; Seyer, Jerome M.

In: Topics in Catalysis, Vol. 4, No. 6, 01.01.1984, p. 479-492.

Research output: Contribution to journalArticle

Mainardi, Carlo L. ; Hibbs, Margaret S. ; Hasty, Karen ; Seyer, Jerome M. / Purification of a Type V Collagen Degrading Metalloproteinase from Rabbit Alveolar Macrophages. In: Topics in Catalysis. 1984 ; Vol. 4, No. 6. pp. 479-492.
@article{311056254ebb4b43b22160bbd48992aa,
title = "Purification of a Type V Collagen Degrading Metalloproteinase from Rabbit Alveolar Macrophages",
abstract = "A proteinase capable of degrading the helical region of native human type V collagen was identified in serum-free culture medium from “in vivo-activated” rabbit alveolar macrophages. This enzyme was purified to homogeneity using a combination of gelfiltration, ion-exchange and affinity chromatography. Analysis of the purified material by gel electrophoresis revealed a single broad band with relative molecular mass of 82,000 daltons Enzyme activity was eluted from the gel in a region corresponding to the stained band. The protein band was also found to stain positively using the periodic acid-Schiff technique indicating that it was glycosylated. Amino acid analysis revealed a composition rich in acidic residues. The enzyme cleaved type V collagen into three large molecular weight doublets consistent with two cleavage sites within the helix but was inactive against native type I collagen. However, when type I or type V collagen was heat-denatured, the enzyme degraded the alpha chains to small molecular weight peptides.",
author = "Mainardi, {Carlo L.} and Hibbs, {Margaret S.} and Karen Hasty and Seyer, {Jerome M.}",
year = "1984",
month = "1",
day = "1",
doi = "10.1016/S0174-173X(84)80014-X",
language = "English (US)",
volume = "4",
pages = "479--492",
journal = "Topics in Catalysis",
number = "6",

}

TY - JOUR

T1 - Purification of a Type V Collagen Degrading Metalloproteinase from Rabbit Alveolar Macrophages

AU - Mainardi, Carlo L.

AU - Hibbs, Margaret S.

AU - Hasty, Karen

AU - Seyer, Jerome M.

PY - 1984/1/1

Y1 - 1984/1/1

N2 - A proteinase capable of degrading the helical region of native human type V collagen was identified in serum-free culture medium from “in vivo-activated” rabbit alveolar macrophages. This enzyme was purified to homogeneity using a combination of gelfiltration, ion-exchange and affinity chromatography. Analysis of the purified material by gel electrophoresis revealed a single broad band with relative molecular mass of 82,000 daltons Enzyme activity was eluted from the gel in a region corresponding to the stained band. The protein band was also found to stain positively using the periodic acid-Schiff technique indicating that it was glycosylated. Amino acid analysis revealed a composition rich in acidic residues. The enzyme cleaved type V collagen into three large molecular weight doublets consistent with two cleavage sites within the helix but was inactive against native type I collagen. However, when type I or type V collagen was heat-denatured, the enzyme degraded the alpha chains to small molecular weight peptides.

AB - A proteinase capable of degrading the helical region of native human type V collagen was identified in serum-free culture medium from “in vivo-activated” rabbit alveolar macrophages. This enzyme was purified to homogeneity using a combination of gelfiltration, ion-exchange and affinity chromatography. Analysis of the purified material by gel electrophoresis revealed a single broad band with relative molecular mass of 82,000 daltons Enzyme activity was eluted from the gel in a region corresponding to the stained band. The protein band was also found to stain positively using the periodic acid-Schiff technique indicating that it was glycosylated. Amino acid analysis revealed a composition rich in acidic residues. The enzyme cleaved type V collagen into three large molecular weight doublets consistent with two cleavage sites within the helix but was inactive against native type I collagen. However, when type I or type V collagen was heat-denatured, the enzyme degraded the alpha chains to small molecular weight peptides.

UR - http://www.scopus.com/inward/record.url?scp=0021732056&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021732056&partnerID=8YFLogxK

U2 - 10.1016/S0174-173X(84)80014-X

DO - 10.1016/S0174-173X(84)80014-X

M3 - Article

VL - 4

SP - 479

EP - 492

JO - Topics in Catalysis

JF - Topics in Catalysis

IS - 6

ER -