# METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN

> **NIH NIH R01** · UNIVERSITY OF KANSAS MEDICAL CENTER · 1991 · $130,432

## Abstract

Connective tissue cells have the ability to synthesize and secrete
collagenase and at least two other matrix metalloproteases MNP-
2 ("gelatinase") and MMP-3 ("proteoglycanase")) that digest
various extracellular matrix macromolecules, but these enzymes
are secreted from the cells as inactive zymogens.  The long-term
objectives of the proposed research are to investigate the
mechanisms of activation of these zymogens and to identify
active enzymes in situ in order to understand their roles in
pathological destruction as well as in normal turnover of the
connective tissue matrix.

Three zymogens (procollagenase, proMMP-2 and proMMP-3) will
be purified from human rheumatoid synovial cells in culture by
various chromatographic techniques.  The primary structure of
proMMP-2 will be deduced from the DNA sequence which is
complementary to proMMP-2 mRNA.  The sequence information
is used to investigate the mechanisms of activation of the
proenzymes at the sequence level.  Various tissue and plasma
proteases and 4-aminophenylmercuric acetate will be tested for
their abilities to activate proMMP-2 and proMMP-3, and their
actions on proenzymes will be characterized by the NH2-terminal
sequence analyses of active enzymes.  The "cascade" hypothesis
for the accelerated activation of these proenzymes will be also
examined by the recombination of zymogens and activated
enzymes.  Enzymic properties of MMP-2 and MMP-3 will be
further investigated by their action on the basement membrane
components such as type IV collagen, laminin and eutactin.  The
substrate specificities of these enzymes are characterized by
their action on reduced, carboxymethylated transferrin.
Attempts will be made to localize active MMP-3 in the tissues
where rapid matrix resorption is occurring.  Antibodies specific to
the propeptide region of proMMP-3 and monoclonal antibodies
specific to the active MMP-3 will be prepared, and used for the
dual localization of both active and zymogen forms of MMP-3 in
human rheumatoid synovium and articular cartilage, and rabbit
postpartum uterus and interleukin 1-treated articular cartilage.
Once the detailed knowledge is obtained about the zymogen
activation, enzyme specificities, and the sites of activation in the
tissue, it may be possible to suggest specific ways to limit the
unwanted proteolysis of the extracellular matrix that occurs in a
variety of connective tissue diseases.

## Key facts

- **NIH application ID:** 3159159
- **Project number:** 5R01AR039189-04
- **Recipient organization:** UNIVERSITY OF KANSAS MEDICAL CENTER
- **SAM.gov UEI:** YXJGGNC5J269
- **Principal Investigator:** HIDEAKI NAGASE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $130,432
- **Award type:** 5
- **Project period:** 1988-02-01 → 1993-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3159159

## Citation

> US National Institutes of Health, RePORTER application 3159159, METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN (5R01AR039189-04). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3159159. Licensed CC0.

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