# HUMAN SKIN MAST CELL GRANULE ENZYMES--SKIN DISEASES

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 1990 · $222,161

## Abstract

The goals of these studies are to isolate and characterize
carboxypeptidase, a newly identified human mast cell secretory
granule protease, and to localize and determine the pathobiologic
role of mast cell proteolytic enzymes in mast cell mediated skin
diseases.  Proposed studies will address: 1) purification and
physicochemical characterization of human skin mast cell
carboxypeptidase; 2) characterization and in vitro reconstitution
of the human skin mast cell secretory granule with respect to
protease and proteoglycan components; 3) investigation of the
biochemical action of purified carboxypeptidase, and enzyme-
proteoglycan complexes on purified proteins, peptides, and
potential cellular substrates in skin; and 4) examination of the
pathobiologic role of human skin mast cell secretory granule
proteases in mast cell-associated cutaneous diseases.  These
studies will add to understanding of the human skin mast cell, and
the role of mast cell secretory granule proteases in mediating
tissue damage in cutaneous disorders.  To accomplish these goals,
human skin mast cell carboxypeptidase, identified by an HPLC
assay system, will be purified from whole skin extracts by affinity
chromatography.  A monospecific, polyclonal
anticarboxypeptidase antibody will be developed.  To understand
regulation of carboxypeptidase expression, mast cells from
various tissues will be assessed for carboxypeptidase activity by
HPLC, and protein by ELISA or RIA, and a human mast cell cDNA
library will be made to identify recombinant clones corresponding
to carboxypeptidase.  To understand their biochemical and
biologic activity, enzyme-proteoglycan complexes from dispersed
human mast cells or reconstituted from purified components, will
be incubated with protein, peptide, and potential cellular
substrates such as fibrinogen, laminin, substance P, angiotensin I,
and cultured human umbilical vein endothelial cells, respectively.
The proteolytic activity of these complexes will be examined in
vitro using skin explants.  The in situ release, localization and
fate of carboxypeptidase will be examined in vitro using skin
explants, and in vivo after intradermal injection of human C5a,
and in a skin chamber model after challenge with allergens.
Finally, the deposition of mast cell granule carboxypeptidase will
be examined in vivo in diseases marked by increased vascular
permeability, i.e.  cold urticaria, and by tissue destruction, i.e.
bullous pemphigoid.

## Key facts

- **NIH application ID:** 3156146
- **Project number:** 5R01AR031901-08
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **SAM.gov UEI:** KMH5K9V7S518
- **Principal Investigator:** BRUCE U WINTROUB
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1990
- **Award amount:** $222,161
- **Award type:** 5
- **Project period:** 1982-07-01 → 1993-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3156146

## Citation

> US National Institutes of Health, RePORTER application 3156146, HUMAN SKIN MAST CELL GRANULE ENZYMES--SKIN DISEASES (5R01AR031901-08). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3156146. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*