Immunodetection of the expression of microsomal proteins encoded by the glucose 6-phosphate transporter gene

Silvia Senesi, Paola Marcolongo, T. Kardon, Giovanna Bucci, Andrey Sukhodub, Ann Burchell, Angelo Benedetti, Rosella Fulceri

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Glucose 6-phosphate transport has been well characterized in liver microsomes. The transport is required for the functioning of the glucose-6-phosphatase enzyme that is situated in the lumen of the hepatic endoplasmic reticulum. The genetic deficiency of the glucose 6-phosphate transport activity causes a severe metabolic disease termed type 1b glycogen storage disease. The cDNA encoding a liver transporter for glucose 6-phosphate was cloned and was found to be mutated in patients suffering from glycogen storage disease 1b. While related mRNAs have been described in liver and other tissues, the encoded protein(s) has not been immunologically characterized yet. In the present study, we report (using antibodies against three different peptides of the predicted amino acid sequence) that a major protein encoded by the glucose 6-phosphate transporter gene is expressed in the endoplasmic reticulum membranes of rat and human liver. The protein has an apparent molecular mass of approx. 33 kDa using SDS/PAGE, but several lines of evidence indicate that its real molecular mass is 46 kDa, as expected. The glucose 6-phosphate transporter protein was also immunodetected in kidney microsomes, but not in microsomes derived from human fibrocytes, rat spleen and lung, and a variety of cell lines. Moreover, little or no expression of the glucose 6-phosphate transporter protein was found in liver microsomes obtained from three glycogen storage disease 1b patients, even bearing mutations that do not directly interfere with protein translation, which can be explained by a (proteasome-mediated) degradation of the mutated transporter.

Original languageEnglish
Pages (from-to)57-62
Number of pages6
JournalBiochemical Journal
Volume389
Issue number1
DOIs
Publication statusPublished - Jul 1 2005

Fingerprint

Glycogen Storage Disease
Liver
Genes
Glycogen
Glucose-6-Phosphate
Liver Microsomes
Microsomes
Proteins
Endoplasmic Reticulum
Molecular mass
Rats
Bearings (structural)
Glucose-6-Phosphatase
Metabolic Diseases
Protein Biosynthesis
Proteasome Endopeptidase Complex
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Spleen
Complementary DNA

Keywords

  • Endoplasmic reticulum
  • Glucose 6-phosphate transporter
  • Glucose-6-phosphatase
  • Glycogen storage disease
  • Liver
  • Microsome

ASJC Scopus subject areas

  • Biochemistry

Cite this

Immunodetection of the expression of microsomal proteins encoded by the glucose 6-phosphate transporter gene. / Senesi, Silvia; Marcolongo, Paola; Kardon, T.; Bucci, Giovanna; Sukhodub, Andrey; Burchell, Ann; Benedetti, Angelo; Fulceri, Rosella.

In: Biochemical Journal, Vol. 389, No. 1, 01.07.2005, p. 57-62.

Research output: Contribution to journalArticle

Senesi, S, Marcolongo, P, Kardon, T, Bucci, G, Sukhodub, A, Burchell, A, Benedetti, A & Fulceri, R 2005, 'Immunodetection of the expression of microsomal proteins encoded by the glucose 6-phosphate transporter gene', Biochemical Journal, vol. 389, no. 1, pp. 57-62. https://doi.org/10.1042/BJ20050213
Senesi, Silvia ; Marcolongo, Paola ; Kardon, T. ; Bucci, Giovanna ; Sukhodub, Andrey ; Burchell, Ann ; Benedetti, Angelo ; Fulceri, Rosella. / Immunodetection of the expression of microsomal proteins encoded by the glucose 6-phosphate transporter gene. In: Biochemical Journal. 2005 ; Vol. 389, No. 1. pp. 57-62.
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