# METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN

> **NIH NIH R01** · UNIVERSITY OF KANSAS MEDICAL CENTER · 1993 · $148,110

## Abstract

The matrix metalloproteinases (MMPs) are a group of zinc enzymes secreted
from connective tissue cells and phagocytes.  They play a central role
in both normal connective tissue remodeling and in accelerated matrix
breakdown associated with various connective tissue diseases such as
arthritis, gingivitis and tissue ulceration, and in tumor cell invasion
and metastasis.  Eight members of the MMP family in humans have been
identified by cDNA cloning and seven of them have been characterized for
their abilities to degrade extracellular matrix macromolecules.  This
includes collagenases (MMP-1 and MMP-8), gelatinases (MMP-2 and MMP-9),
stromelysins (MMP-3 and MMP-10) and a 28-kDa metalloproteinase/Pump-
1/matrilysin (MMP-7).  While the synthesis of many MMPs is
transcriptionally regulated by various cytokines and growth factors, all
of them are secreted from the cells as inactive zymogens (proMMPs).  The
latter event emphasizes that the extracellular activation processes of
the zymogens and their interaction with matrix substrates and endogenous
inhibitors are also important regulatory steps in tissue catabolism.  The
major objective of this proposal is to investigate the activation and the
catalytic mechanisms of MMPs in order to understand the precise
biological and pathological roles of these enzymes in connective tissue
destruction.

The catalytic mechanism of MMPs will be investigated using the 28-kDa
MMP-3 (stromelysin 1) as a prototype.  this will be approached by
identification of the side chains involved in binding to tissue inhibitor
of metalloproteinases 1 (TIMP-1) and in reaction with alpha2-
macroglobulin by a competitive binding "label selection" method.  The
predicted residues by this study will then be tested by site-directed
mutagenesis.  In parallel, X-ray crystallographic studies will be
conducted in collaboration with Dr. W. Bode at Max-Planck-Institut in
Munich.  The structural basis for MMP-1 (interstitial collagenase) to
express collagenolytic activity will be investigated by homolog-scanning
and alanine-scanning mutageneses and by a "differential trace labeling"
method.  The zymogen activation studies will focus on the kinetic
activation analyses of proMMP-2 (72-kDa gelatinase) and proMMP-9 (92-kDa
gelatinase) when they are complexed with TIMP-2 and TIMP-1, respectively.
Once the detailed knowledge is obtained about the structural and
functional relationship of different MMPs to account for their substrate
specificities and their catalytic mechanisms, it will be possible to
design synthetic inhibitors that can manipulate the unwanted proteolysis
of the extracellular matrix that occurs in a variety of connective tissue
diseases.

## Key facts

- **NIH application ID:** 3159156
- **Project number:** 2R01AR039189-06A1
- **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:** 1993
- **Award amount:** $148,110
- **Award type:** 2
- **Project period:** 1988-02-01 → 1997-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3159156

## Citation

> US National Institutes of Health, RePORTER application 3159156, METALLOPROTEASES IN CONNECTIVE TISSUE MATRIX BREAKDOWN (2R01AR039189-06A1). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3159156. Licensed CC0.

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