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PMID: 8755516 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Expression cloning of the Golgi CMP-sialic acid transporter.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 15 ·1996-07-23 ·Pages 7572-6

Eckhardt M, Mühlenhoff M, Bethe A, Gerardy-Schahn R

Abstract

Translocation of nucleotide sugars across the membrane of the Golgi apparatus is a prerequisite for the synthesis of complex carbohydrate structures. While specific transport systems for different nucleotide sugars have been identified biochemically in isolated microsomes and Golgi vesicles, none of these transport proteins has been characterized at the molecular level. Chinese hamster ovary (CHO) mutants of the complementation group Lec2 exhibit a strong reduction in sialylation of glycoproteins and glycolipids due to a defect in the CMP-sialic acid transport system. By complementation cloning in the mutant 6B2, belonging to the Lec2 complementation group, we were able to isolate a cDNA encoding the putative murine Golgi CMP-sialic acid transporter. The cloned cDNA encodes a highly hydrophobic, multiple membrane spanning protein of 36.4 kDa, with structural similarity to the recently cloned ammonium transporters. Transfection of a hemagglutinin-tagged fusion protein into the mutant 6B2 led to Golgi localization of the hemagglutinin epitope. Our results, together with the observation that the cloned gene shares structural similarities to other recently cloned transporter proteins, strongly suggest that the isolated cDNA encodes the CMP-sialic acid transporter.

MeSH Terms
Amino Acid Sequence Animals Base Sequence CHO Cells Carrier Proteins/biosynthesis,chemistry,metabolism Cloning, Molecular Cricetinae Cytidine Monophosphate N-Acetylneuraminic Acid/metabolism Genetic Complementation Test Glycolipids/biosynthesis Glycoproteins/biosynthesis Golgi Apparatus/metabolism Hemagglutinins/biosynthesis Membrane Proteins/biosynthesis,chemistry,metabolism Mice Molecular Sequence Data Molecular Weight Nucleotide Transport Proteins Recombinant Fusion Proteins/biosynthesis
Chemicals
Carrier Proteins Glycolipids Glycoproteins Hemagglutinins Membrane Proteins Nucleotide Transport Proteins Recombinant Fusion Proteins Slc35a1 protein, mouse Cytidine Monophosphate N-Acetylneuraminic Acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Eckhardt M
Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, Germany.
Mühlenhoff M
Bethe A
Gerardy-Schahn R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.15.7572
Published
1996-07-23
Pages
7572-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC38787
Subset
IM
Databases
GENBANK
Z71268
External Links
PubMed source DOI: 10.1073/pnas.93.15.7572 PMC Full Text View journal details
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