Proteomics

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Sac1 interaction proteomics of ALFA Sac1 and mutants

ABSTRACT: Sphingolipids are structural membrane components that also function in cellular stress responses. The serine palmitoyl-transferase (SPT) catalyzes the rate limiting step in sphingolipid biogenesis. Its activity is tightly regulated through multiple bind-ing partners, including Tsc3, Orm proteins, ceramides, and the phosphatidylinositol-4-phosphate (PI4P) phosphatase Sac1. The structural organization and regulatory mechanisms of this complex are not yet understood. Here, we report the high-resolution cryo-EM structures of the yeast SPT in complex with Tsc3 and Orm1 (SPOT) as dimers and monomers and a monomeric complex further carrying Sac1 (SPOTS). In all complexes, the tight interaction of the down-stream metabolite ceramide and Orm1 reveals the ceramide dependent inhibition. Additionally, observation of ceramide and ergosterol binding suggests a co-regulation of sphingolipid biogenesis and sterol metabolism within the SPOTS com-plex.

INSTRUMENT(S): Bruker Daltonics timsTOF series

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Florian Fröhlich  

LAB HEAD: Florian Fröhlich

PROVIDER: PXD044586 | Pride | 2023-10-24

REPOSITORIES: Pride

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Dataset's files

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Action DRS
CTRL1_2uL1h_681.d.zip Other
CTRL2_2uL1h_682.d.zip Other
CTRL3_2uL1h_683.d.zip Other
Sac1D573_1_2uL1h_685.d.zip Other
Sac1D573_2_2uL1h_686.d.zip Other
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Publications

Sphingolipids are structural membrane components that also function in cellular stress responses. The serine palmitoyltransferase (SPT) catalyzes the rate-limiting step in sphingolipid biogenesis. Its activity is tightly regulated through multiple binding partners, including Tsc3, Orm proteins, ceramides, and the phosphatidylinositol-4-phosphate (PI4P) phosphatase Sac1. The structural organization and regulatory mechanisms of this complex are not yet understood. Here, we report the high-resoluti  ...[more]

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