Identification of centaurin-alpha 2: a phosphatidylinositide-binding protein present in fat, heart and skeletal muscle

P Whitley, A M Gibbard, F Koumanov, S Oldfield, E E Kilgour, G D Prestwich, G D Holman

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Abstract

We describe here the cloning, expression and characterisation of centaurin-alpha2 from a rat adipocyte cDNA library. The centaurin-alpha2 cDNA contains an open reading frame, which codes for a protein of 376 amino acids with predicted mass of 43.5 kDa. Centaurin-alpha2 shares 51-59% identity with centaurin-alpha1 proteins and has the same domain organisation, consisting of a predicted N-terminal ArfGAP domain followed by two successive pleckstrin homology domains. Despite the sequence similarity, there are a number of notable differences between the previously characterised centaurin-alpha1 proteins and the newly described centaurin-alpha2: (i) in vitro lipid binding experiments with centaurin-alpha2 do not reveal the same selectivity for phosphatidylinositol 3,4,5-trisphosphate over phosphatidylinositol 4,5-bisphosphate that has been shown for centaurin-alpha1; (H) unlike centaurin-alpha1 which is expressed mainly in the brain, centaurin-alpha2 has a broad tissue distribution, being particularly abundant in fat, heart and skeletal muscle; (iii) in contrast to centaurin-alpha1 which is found in both membrane and cytosolic fractions, endogenous centaurin-alpha2 is exclusively present in the dense membrane fractions of cell extracts, suggesting a constitutive membrane association. Insulin stimulation, which stimulates phosphatidylinositol 3,4,5-trisphosphate production, does not alter the subcellular distribution of centaurin-alpha2 between adipocyte membrane fractions. This observation is consistent with the lack of specificity of centaurin-alpha2 for phosphatidylinositol 3,4,5-trisphosphate over phosphatidylinositol 4,5-bisphosphate.
Original languageEnglish
Pages (from-to)222-230
Number of pages9
JournalEuropean Journal of Cell Biology
Volume81
Issue number4
DOIs
Publication statusPublished - Apr 2002

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Myocardium
Carrier Proteins
Skeletal Muscle
Fats
Membranes
Phosphatidylinositols
Adipocytes
Proteins
Tissue Distribution
Cell Extracts
Gene Library
Open Reading Frames
Organism Cloning
Complementary DNA
Insulin
Lipids
Amino Acids
Brain
phosphatidylinositol 3,4,5-triphosphate

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Identification of centaurin-alpha 2: a phosphatidylinositide-binding protein present in fat, heart and skeletal muscle. / Whitley, P; Gibbard, A M; Koumanov, F; Oldfield, S; Kilgour, E E; Prestwich, G D; Holman, G D.

In: European Journal of Cell Biology, Vol. 81, No. 4, 04.2002, p. 222-230.

Research output: Contribution to journalArticle

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