Functional expression of the murine Golgi CMP-sialic acid transporter in Saccharomyces cerevisiae

We have functionally expressed the murine Golgi putative CMP-sialic acid transporter in Saccharomyces cerevisiae . Using a galactose-inducible expression system, S. cerevisiae vesicles were able to transport CMP-sialic acid. Transport was dependent on galactose induction and was temperature-dependen...

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Bibliographic Details
Published in:The Journal of biological chemistry 1997-05, Vol.272 (19), p.12616-12619
Main Authors: Berninsone, P, Eckhardt, M, Gerardy-Schahn, R, Hirschberg, C.B
Format: Article
Language:English
Subjects:
ADN
AMINO SUGARS
AMINOAZUCARES
AMINOSUCRE
Animals
Carrier Proteins - metabolism
Cells, Cultured
CITOSINA
COMPLEMENTARY DNA
CYTIDINE MONOPHOSPHATE
Cytidine Monophosphate N-Acetylneuraminic Acid - metabolism
CYTOSINE
DNA
EXPRESION GENICA
EXPRESSION DES GENES
GALACTOSA
GALACTOSE
GENE EXPRESSION
GENETIC VECTORS
Golgi Apparatus - metabolism
Kinetics
Membrane Proteins - metabolism
Mice
NUCLEOTIDE
Nucleotide Transport Proteins
NUCLEOTIDES
NUCLEOTIDOS
PIRIMIDINAS
PROTEINAS
PROTEINE
PROTEINS
PYRIMIDINE
PYRIMIDINES
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - metabolism
TRANSPORT PROCESSES
TRANSPORT PROTEINS
UDP
VECTEUR GENETIQUE
VECTORES GENETICOS
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Description
Summary:We have functionally expressed the murine Golgi putative CMP-sialic acid transporter in Saccharomyces cerevisiae . Using a galactose-inducible expression system, S. cerevisiae vesicles were able to transport CMP-sialic acid. Transport was dependent on galactose induction and was temperature-dependent and saturable with an apparent K m of 2.9 μ m . Transport was inhibited by CMP, and upon vesicle disruption with Triton X-100 parameters were very similar to the previously described CMP-sialic acid transport characteristics observed with mammalian Golgi vesicles. CMP-sialic acid transport induction was specific as no transport of UDP-galactose was observed even though the latter putative transporter has a high degree of amino acid sequence identity with the CMP-sialic acid transporter. Together, the above results demonstrate that the previously described cDNA encoding the putative CMP-sialic acid transporter encodes the transporter protein per se and suggests that this heterologous expression system may be used for further structural and functional studies of other Golgi membrane transporter proteins.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.272.19.12616