Seipin is a discrete homooligomer

Derk Binns, Sungkyung Lee, Christopher L. Hilton, Qiu Xing Jiang, Joel M. Goodman

Research output: Contribution to journalArticle

78 Scopus citations

Abstract

Seipin is a transmembrane protein that resides in the endoplasmic reticulum and concentrates at junctions between the ER and cytosolic lipid droplets. Mutations in the human seipin gene, including the missense mutation A212P, lead to congenital generalized lipodystrophy (CGL), characterized by the lack of normal adipose tissue and accumulation of fat in liver and muscles. In both yeast and CGL patient fibroblasts, seipin is required for normal lipid droplet morphology; in its absence droplets appear to bud abnormally from the ER. Here we report the first purification and physical characterization of seipin. Yeast seipin is in a large discrete protein complex. Affinity purification demonstrated that seipin is the main if not exclusive protein in the complex. Detergent sucrose gradients in H2O, and D2O and gel filtration were used to determine the size of the seipin complex and account for detergent binding. Both seipin-myc13 (seipin fused to 13 tandem copies of the myc epitope) expressed from the endogenous promoter and overexpressed seipin-mCherry form ∼500 kDa proteins consisting of about 9 copies of seipin. The yeast orthologue of the human A212P allele forms only smaller complexes and is unstable; we hypothesize that this accounts for its null phenotype in humans. Seipin appears as a toroid by negative staining electron microscopy. We speculate that seipin plays at least a structural role in organizing droplets or in communication between droplets and ER.

Original languageEnglish (US)
Pages (from-to)10747-10755
Number of pages9
JournalBiochemistry
Volume49
Issue number50
DOIs
StatePublished - Dec 21 2010

ASJC Scopus subject areas

  • Biochemistry

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    Binns, D., Lee, S., Hilton, C. L., Jiang, Q. X., & Goodman, J. M. (2010). Seipin is a discrete homooligomer. Biochemistry, 49(50), 10747-10755. https://doi.org/10.1021/bi1013003