# STRUCTURE AND ORGANIZATION OF BASEMENT MEMBRANES

> **NIH NIH R01** · UNIV OF MED/DENT NJ-R W JOHNSON MED SCH · 1987 · $123,362

## Abstract

Basement membranes are extracellular matrices located between epithelial or
endothelial cells and underlying connective tissues.  They function as
selective barriers and are of critical importance in maintaining normal
tissue function and integrity.  The major components of basement membranes
are proteins designated Type IV collagens, non-collagenous glycoproteins
(laminin) and proteoglycans.  The goal of the research proposed here is to
determine the complete covalent structure of human Type IV collagens which
are thought to be the main structural component of basement membranes.

To accomplish this goal we will use two different approaches:

(a) We will synthesize complementary DNA (cDNA) to Type IV
collagen-specific mRNA isolated from a human tumor cell line (American Type
Culture Collection, HT-1080 Cells).  The cDNA will be inserted into pBR322
and grown in E. coli strain HB101.  DNA sequencing will be carried out and
the sequence will be analyzed to yield an amino acid sequence.

(b) We will isolate intact Type IV collagens from cultured HT-1080 cells
and collagen fragments solubilized by limited pepsin digestion from human
placental tissue.  Post-translational modifications such as hydroxylation
of prolyl and lysyl residues, glycosylation of hydroxylysyl and asparaginyl
residues, location of disulfide bonds and cleavage sites of processing
enzymes will be determined by direct protein sequencing.

The long term objective of this research is to understand the structures
and its relation to the function of basement membranes in normal and
diseased states.  As a first step in this direction, we will determine the
complete covalent structure of basement membrane collagens using a dual
approach:  protein and DNA sequencing.

Together these complementary approaches provide the most effective and
rapid means of achieving the research goal.  This work will provide a solid
basis for further structural studies and will provide specific probes for
investigating changes in basement membranes caused by pathological
conditions such as are found in diabetes.

## Key facts

- **NIH application ID:** 3155233
- **Project number:** 5R01AR022051-09
- **Recipient organization:** UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
- **Principal Investigator:** CHARLES D. BOYD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $123,362
- **Award type:** 5
- **Project period:** 1978-07-01 → 1988-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3155233

## Citation

> US National Institutes of Health, RePORTER application 3155233, STRUCTURE AND ORGANIZATION OF BASEMENT MEMBRANES (5R01AR022051-09). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3155233. Licensed CC0.

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