Pantophysin is a phosphoprotein component of adipocyte transport vesicles and associates with GLUT4-containing vesicles

Cydney C. Brooks, Philipp E. Scherer, Kelly Cleveland, Jennifer L. Whittemore, Harvey F. Lodish, Bentley Cheatham

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Abstract

Pantophysin, a protein related to the neuroendocrin-especific synaptophysin, recently has been identified in non-neuronal tissues. In the present study, Northern blots showed that pantophysin mRNA was abundant in adipose tissue and increased during adipogenesis of 3T3-L1 cells. Immunoblot analysis of subcellular fractions showed pantophysin present exclusively in membrane fractions and relatively evenly distributed in the plasma membrane and internal membrane fractions. Sucrose gradient ultracentrifugation demonstrated that pantophysin and GLUT4 exhibited overlapping distribution profiles. Furthermore, immunopurified GLUT4 vesicles contained pantophysin, and both GLUT4 and pantophysin were depleted from this vesicle population following treatment with insulin. Additionally, a subpopulation of immunopurified pantophysin vesicles contained insulin-responsive GLUT4. Consistent with the interaction of synaptophysin with vesicle-associated membrane protein 2 in neuroendocrine tissues, pantophysin associated with vesicle-associated membrane protein 2 in adipocytes. Furthermore, in [32P]orthophosphate-labeled cells, pantophysin was phosphorylated in the basal state. This phosphorylation was unchanged in response to insulin; however, insulin stimulated the phosphorylation of a 77-kDa protein associated with α-pantophysin immunoprecipitates. Although the functional role of pantophysin in vesicle trafficking is unclear, its presence on GLUT4 vesicles is consistent with the emerging role of soluble N-ethylmaleimide- sensitive protein receptor (SNARE) factor complex and related proteins in regulated vesicle transport in adipocytes. In addition, pantophysin may provide a marker for the analysis of other vesicles in adipocytes.

Original languageEnglish (US)
Pages (from-to)2029-2036
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number3
DOIs
StatePublished - Jan 21 2000

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Transport Vesicles
Phosphoproteins
Adipocytes
Vesicle-Associated Membrane Protein 2
Insulin
Synaptophysin
Phosphorylation
Tissue
N-Ethylmaleimide-Sensitive Proteins
3T3-L1 Cells
Membranes
SNARE Proteins
Adipogenesis
Proteins
Subcellular Fractions
Ultracentrifugation
Cell membranes
Northern Blotting
Sucrose
Adipose Tissue

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pantophysin is a phosphoprotein component of adipocyte transport vesicles and associates with GLUT4-containing vesicles. / Brooks, Cydney C.; Scherer, Philipp E.; Cleveland, Kelly; Whittemore, Jennifer L.; Lodish, Harvey F.; Cheatham, Bentley.

In: Journal of Biological Chemistry, Vol. 275, No. 3, 21.01.2000, p. 2029-2036.

Research output: Contribution to journalArticle

Brooks, Cydney C. ; Scherer, Philipp E. ; Cleveland, Kelly ; Whittemore, Jennifer L. ; Lodish, Harvey F. ; Cheatham, Bentley. / Pantophysin is a phosphoprotein component of adipocyte transport vesicles and associates with GLUT4-containing vesicles. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 3. pp. 2029-2036.
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