Recombinant Saccharomyces cerevisiae Mannosyl phosphorylinositol ceramide synthase CSH1 (CSH1)
Description
Introduction to Recombinant Saccharomyces cerevisiae Mannosyl Phosphorylinositol Ceramide Synthase CSH1 (CSH1)
Recombinant Saccharomyces cerevisiae Mannosyl phosphorylinositol ceramide synthase CSH1 (CSH1) is an enzyme involved in the synthesis of complex sphingolipids, specifically mannosylinositol phosphorylceramides (MIPCs), in the yeast Saccharomyces cerevisiae. This enzyme is crucial for maintaining cell wall integrity and plays a role in the coordination of sphingolipid metabolism with other cellular processes.
Function and Role of CSH1
CSH1, along with its homolog SUR1, is responsible for synthesizing MIPCs in the Golgi apparatus of S. cerevisiae. These sphingolipids are essential for cell wall integrity and interact with ergosterol to maintain the structural and functional integrity of the cell wall . The deletion of CSH1 and SUR1 genes can lead to increased phosphorylation of the mitogen-activated protein kinase Slt2, indicating a defect in cell wall integrity .
Structure and Modifications
CSH1 contains N-glycans, which are important for its enzymatic activity and stability. Specifically, CSH1 has N-glycans on Asn-51 and Asn-247. The N-glycan on Asn-51 exhibits a unique mannan-like structure, which is distinct from the typical core-type N-glycans found in intracellular proteins . These modifications play a critical role in substrate recognition and enzyme stability.
Recombinant Production
Recombinant CSH1 can be produced in various hosts, including E. coli, yeast, baculovirus, and mammalian cells, with a purity of ≥85% as determined by SDS-PAGE . This recombinant enzyme is useful for studying the biochemical properties of CSH1 and its role in sphingolipid synthesis.
Research Findings
Recent studies have highlighted the importance of CSH1 in sphingolipid metabolism and cell wall integrity. The interaction between MIPCs synthesized by CSH1 and ergosterol is crucial for maintaining cell wall function . Additionally, the unique N-glycan structures on CSH1 influence its enzymatic activity and stability, suggesting a complex regulatory mechanism for sphingolipid synthesis .
Data Table: Characteristics of Recombinant CSH1
| Characteristic | Description |
|---|---|
| Gene Name | CSH1 |
| Hosts | E. coli, Yeast, Baculovirus, Mammalian Cells |
| Purity | ≥85% (SDS-PAGE) |
| N-Glycosylation Sites | Asn-51, Asn-247 |
| Function | Synthesis of mannosylinositol phosphorylceramides (MIPCs) |
Product Specs
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Target Background
Function: Involved in the synthesis of mannosyl phosphorylinositol ceramide. Catalyzes the addition of mannose to phosphorylinositol ceramide.
KEGG: sce:YBR161W
STRING: 4932.YBR161W
