# ASPARAGINE-LINKED CARBOHYDRATES

> **NIH NIH R01** · UNIVERSITY OF MIAMI SCHOOL OF MEDICINE · 1989 · $19,766

## Abstract

Our research for the next year involves the completion of our analysis of
the asparagine-linked oligosaccharides of BHK and polyoma-transformed BHK
cells by sequential lectin-affinity chromatography (SLAC) and methylation
analyses.  Two lectins, E-phytohemagglutinin and one from Datura stamonium,
will be added to the analysis of the glycopeptides.  Next, each of the 29
L-PHA-resistant PyBHK clones will be screened for its ability not to bind
labeled L-PHA using the procedure we have developed for the parental PyBHK
and the BHK cells.  Any clone which can be isolated that is L-PHA- will
be radiolabeled with [2-3H]-mannose, and its Asnlinked glycopeptides
analyzed by SLAC and compared to those from the parental PyBHK cells.  The
next series of experiments involves the production of tumors in hamsters by
injection of PyBHK cells and their lectin-resistant variants.  We will be
able to compare the Asn-linked glycopeptides of the tumors by radiolabeling
in organ culture and then SLAC analysis.  We wish to know if a particular
oligosaccharide phenotype is selected for in vivo.  We will finish the
analysis of the glycosyltransferases that participate in the synthesis of
the Asn-linked oligosaccharides.  We will determine if the specific
activities of these enzymes change upon transformation and will attempt to
explain the differences in the oligosaccharide structures between the two
cell types in terms of enzyme activity differences.  The final aim will be
to finish the examination of the various glycoproteins on the surfaces of
the BHK, PyBHK, and variant cells which express the highly branched
structures and determine if the increase in branched structures occurs on
all or only a few of these glycoproteins.  This analysis, coupled with the
results of the glycosyltransferase activity comparisons, should allow us to
focus on the immediate cause in the changes in oligosaccharide branching
patterns.  (A)

## Key facts

- **NIH application ID:** 3172985
- **Project number:** 3R01CA035377-06S1
- **Recipient organization:** UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
- **SAM.gov UEI:** F8THLJQSAF93
- **Principal Investigator:** J. Michael Pierce
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $19,766
- **Award type:** 3
- **Project period:** 1983-08-01 → 1989-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3172985

## Citation

> US National Institutes of Health, RePORTER application 3172985, ASPARAGINE-LINKED CARBOHYDRATES (3R01CA035377-06S1). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3172985. Licensed CC0.

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