Abstract

The objective of the proposed research is to investigate the mechanism of action of collagenases, which are unique in their ability to catalyze the degradation of collagen under physiological conditions. Collagen, the major structural protein of connective tissue, is highly resistant to attack by all known digestive and intracellular enzymes. Hence, the action of collagenases is essential to all processes involving the degradation or remodeling of connective tissue. Collagenases, which are believed to be zinc metalloenzymes, will be isolated from various sources according to published procedures and by affinity chromatographic methods to be developed. The enzymes obtained will be characterized with respect to their homogeneity, metal content, amino acid composition, molecular weight and subunit structure. The role of the metal atom(s) in catalysis will be investigated by examining the effect of metal substitutions on enzymatic activity. The substrate specificities of these enzymes toward native collagen (Types I-IV) and towards a series of synthetic peptides will be examined in detail. Chemical modification reactions will be carried out to identify essential amino acid residues. This research should promote a greater understanding of the role of collagenases in mammalian connective tissue metabolism.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM027939-06S1
Application #
3275185
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1980-04-01
Project End
1986-11-30
Budget Start
1986-04-01
Budget End
1986-11-30
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Florida State University
Department
Type
Schools of Arts and Sciences
DUNS #
020520466
City
Tallahassee
State
FL
Country
United States
Zip Code
32306

  Publications

Holman, C M; Kan, C C; Gehring, M R et al. (1999) Role of His-224 in the anomalous pH dependence of human stromelysin-1. Biochemistry 38:677-81
Welch, A R; Holman, C M; Huber, M et al. (1996) Understanding the P1' specificity of the matrix metalloproteinases: effect of S1' pocket mutations in matrilysin and stromelysin-1. Biochemistry 35:10103-9
Springman, E B; Nagase, H; Birkedal-Hansen, H et al. (1995) Zinc content and function in human fibroblast collagenase. Biochemistry 34:15713-20
Welch, A R; Holman, C M; Browner, M F et al. (1995) Purification of human matrilysin produced in Escherichia coli and characterization using a new optimized fluorogenic peptide substrate. Arch Biochem Biophys 324:59-64
Netzel-Arnett, S; Salari, A; Goli, U B et al. (1994) Evidence for a triple helix recognition site in the hemopexin-like domains of human fibroblast and neutrophil interstitial collagenases. Ann N Y Acad Sci 732:22-30
Netzel-Arnett, S; Sang, Q X; Moore, W G et al. (1993) Comparative sequence specificities of human 72- and 92-kDa gelatinases (type IV collagenases) and PUMP (matrilysin). Biochemistry 32:6427-32
Angleton, E L; Nagase, H; Birkedal-Hansen, H et al. (1992) Fluorogenic peptide substrates optimized for five human matrix metalloproteinases. Matrix Suppl 1:89-90
French, M F; Bhown, A; Van Wart, H E (1992) Identification of Clostridium histolyticum collagenase hyperreactive sites in type I, II, and III collagens: lack of correlation with local triple helical stability. J Protein Chem 11:83-97
Mookhtiar, K A; Van Wart, H E (1992) Clostridium histolyticum collagenases: a new look at some old enzymes. Matrix Suppl 1:116-26
Devarajan, P; Johnston, J J; Ginsberg, S S et al. (1992) Structure and expression of neutrophil gelatinase cDNA. Identity with type IV collagenase from HT1080 cells. J Biol Chem 267:25228-32

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