# PROTEINS OF THE HEPATOMA CELL PLASMA MEMBRANE

> **NIH NIH R01** · STATE UNIVERSITY OF NEW YORK AT BUFFALO · 1991 · $15,628

## Abstract

The overall objective of the proposed research is to identify by a variety
of methods, including external labeling, immunological, cell fractionation
and cytochemical the proteins and the glycoproteins that are externally
oriented in the plasma membrane of hepatoma cells and primary cultures of
rat hepatocytes.  Lactoperoxidase catalyzed iodination and tritiated sodium
borohydride reduction of cells in situ are used to label externally
accessible proteins and glycoproteins.  These proteins then are resolved by
two-dimensional polyacrylamide gel electrophoresis in the presence of
detergents.  A number of these proteins are being purified and antibodies
both conventional and monoclonal are being prepared to them.  Cell
fractionation in combination with immunocytochemical localization studies
at both the light and electron microscopic level then are used to assign
each of the proteins to the domain of the plasma membrane in which it
resides.  Antibodies to different proteins in different domains of the
hepatocyte plasma membrane next are used to isolate the different domains
of the membrane in pure form using novel methodology not based on
ultracentrifugation.  The composition of the other proteins in the purified
domain is being established by two-dimensional polyacrylamide gel
electrophoretic analyses.  The turnover behavior of each of the proteins in
each of the domains is being assessed utilizing the dual isotopic labeling
procedures that have already been developed in our laboratory.  The time
course of appearance during development of domain specific proteins also is
being examined.  The goal of these studies is to establish if the domain is
the unit for membrane turnover in this complex cell type and to determine
which of the three most plausible mechanisms; lysosomal fusion,
nonlysosomal proteases and/or shedding, is involved in the regulation of
plasma membrane protein concentration in hepatocytes.  As a corollary to
this primary goal studies are being conducted to follow the pathway of
biogenesis of specific proteins to their domain of residence in both the
plasma membrane of the differentiated hepatocyte and the hepatocyte during
development.

## Key facts

- **NIH application ID:** 3177033
- **Project number:** 3R01CA038773-07S1
- **Recipient organization:** STATE UNIVERSITY OF NEW YORK AT BUFFALO
- **SAM.gov UEI:** LMCJKRFW5R81
- **Principal Investigator:** DARRELL J DOYLE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $15,628
- **Award type:** 3
- **Project period:** 1984-05-01 → 1992-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3177033

## Citation

> US National Institutes of Health, RePORTER application 3177033, PROTEINS OF THE HEPATOMA CELL PLASMA MEMBRANE (3R01CA038773-07S1). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3177033. Licensed CC0.

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