Abstract

A study is proprosed of the function and regulation of neutral secretory proteases in keratinocyte-mediated interstitial collagen and basement-membrane degradation. (1) The ability of live keratinocytes to degrade interstitial type I and III collagen fibrils in culture will be measured and the specific and coordinate role of the component proteases of the collagenolytic complex: a 65K collagenase, a 110K gelatinase, two (70K and 48K) plasminogen activators and exogenous plasminogen will be analyzed by immune-inhibition. A search will be made for additional components of the collagenolytic complex such as specific activators of procollagenase and plasminogen proactivators and the regulatory role of plasma and endogenous secretory protease inhibitors, including epidermal inhibitors of the fibrinolytic and collagenolytic systems, will be analyzed. (2) The ability of keratinoyctes to degrade intact basement membranes will be investigated. The component proteases of the type IV collagen, laminin, fibronectin and heparan sulfate proteoglycan degrading systems will be identified and the specific function of the type IV-collagen-degrading protease system will be analyzed by specific immune inhibition. (3) The expression of proteolytic competence as a function of basal cell activation will be investigated. (a) The inverse relationship between secretion of specific proteases and terminal differentiation induced by anchorage deprivation in vitro will be established and the distribution and position of secretor cells in surface-attached primary explants and in clonal colonies will be correlated with that of differentiation markers. (b) The synthesis of proteases by activated basal cells induced by excision wounding in vivo will be studied by immunofluorescence and correlated with migration, confluence and stratification during the healing process. (c) The effect on interstitial collagen and basement membrane degradation of agents which modulate expression of activated and differentiated cellular states such as retinoids, glucocorticoids, inflammatory agents and cyclic nucleotides will be analyzed.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE006028-07
Application #
3219762
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1984-01-01
Project End
1993-12-31
Budget Start
1990-01-01
Budget End
1993-12-31
Support Year
7
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
University of Alabama Birmingham
Department
Type
Schools of Dentistry
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294

  Publications

DeCarlo, A A; Grenett, H E; Harber, G J et al. (1998) Induction of matrix metalloproteinases and a collagen-degrading phenotype in fibroblasts and epithelial cells by secreted Porphyromonas gingivalis proteinase. J Periodontal Res 33:408-20
DeCarlo Jr, A A; Windsor, L J; Bodden, M K et al. (1997) Activation and novel processing of matrix metalloproteinases by a thiol-proteinase from the oral anaerobe Porphyromonas gingivalis. J Dent Res 76:1260-70
Springman, E B; Nagase, H; Birkedal-Hansen, H et al. (1995) Zinc content and function in human fibroblast collagenase. Biochemistry 34:15713-20
Windsor, L J; Bodden, M K; Birkedal-Hansen, B et al. (1994) Mutational analysis of residues in and around the active site of human fibroblast-type collagenase. J Biol Chem 269:26201-7
Birkedal-Hansen, H; Moore, W G; Bodden, M K et al. (1993) Matrix metalloproteinases: a review. Crit Rev Oral Biol Med 4:197-250
Birkedal-Hansen, H (1993) Role of matrix metalloproteinases in human periodontal diseases. J Periodontol 64:474-84
Windsor, L J; Grenett, H; Birkedal-Hansen, B et al. (1993) Cell type-specific regulation of SL-1 and SL-2 genes. Induction of the SL-2 gene but not the SL-1 gene by human keratinocytes in response to cytokines and phorbolesters. J Biol Chem 268:17341-7
Lyons, J G; Birkedal-Hansen, B; Pierson, M C et al. (1993) Interleukin-1 beta and transforming growth factor-alpha/epidermal growth factor induce expression of M(r) 95,000 type IV collagenase/gelatinase and interstitial fibroblast-type collagenase by rat mucosal keratinocytes. J Biol Chem 268:19143-51
Sottrup-Jensen, L; Birkedal-Hansen, H (1992) Localization of cleavage sites for human fibroblast collagenase in the bait region of five mammalian alpha-macroglobulins. Matrix Suppl 1:263-8
Lyons, J G; Birkedal-Hansen, B; Moore, W G et al. (1992) Mr 95,000 metalloproteinase produced by rat mammary epithelial cells. Matrix Suppl 1:85-6

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