Detection of biosynthetic intermediates in proteoglycan-deficient mutants of Chinese hamster ovary cells
Chinese hamster ovary cell mutants lacking xylosyltransferase or galactosyltransferase I do not synthesize mature proteoglycans. The authors predicted that the mutants would accumulate biosynthetic intermediates upstream from the block imposed by mutation. Using the fusogenic properties of vesicular stomatitis virus, the authors fused monolayers composed of galactosyltransferase I-deficient cells with virus-infected xylosyltransferase-deficient cells. Immediately following fusion the cells were pulse-labelled with /sup 35/SO/sub 4/ for one hour. Quantification of radioactive products showed that the mutants contained biosynthetically active intermediates that proceeded to mature glycosaminoglycans. The production of glycosaminoglycan was dependent on fusion, and fusion of each mutant to itself did not result in radioactive product. Analysis of the newly made glycosaminoglycans through HPLC anion-exchange chromatography showed that the fused cells synthesized heparan sulfate and chondroitin sulfate in about the same proportion as wildtype cells. These findings suggest that the mutants accumulate precursors to both families of proteoglycans. They also found that progeny virus from infected CHO cells contain proteoglycans, presumably derived from the plasma membrane. This observation suggests that the virus can be used to isolate intermediates accumulating in the mutants.
- Research Organization:
- Univ. of Alabama, Birmingham
- OSTI ID:
- 6074640
- Report Number(s):
- CONF-870644-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLUCOPROTEINS
BIOSYNTHESIS
CHO CELLS
ION EXCHANGE CHROMATOGRAPHY
LIQUID COLUMN CHROMATOGRAPHY
MUTANTS
SULFATES
SULFUR 35
TRACER TECHNIQUES
VIRUSES
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOHYDRATES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
EVEN-ODD NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PARASITES
PROTEINS
RADIOISOTOPES
SACCHARIDES
SEPARATION PROCESSES
SULFUR COMPOUNDS
SULFUR ISOTOPES
SYNTHESIS
550201* - Biochemistry- Tracer Techniques