Cloned (Comment) | Organism |
---|---|
co-overexpression of His-tagged Erf2 with fused FLAG-tagged Erf4 in strain RJY1827, expression of FLAG-tagged Erf2 in strain RJY | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a fusion protein that places the 58 amino acids of Erf2 onto the C-terminus of Pfa4, another DHHC enzyme that resides in the endoplasmic reticulum membrane, the modification highly reduces the half-life of the engineered protein | Saccharomyces cerevisiae |
General Stability | Organism |
---|---|
Erf4 and the C-terminal 58 amino acids of Erf2 are required for Erf2 stability | Saccharomyces cerevisiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
endoplasmic reticulum | the Erf2-Erf4 complex localizes to the endoplasmic reticulum where it palmitoylates Ras. Role of the quality control system in degrading Erf2 molecules that are not in complex with Erf4 | Saccharomyces cerevisiae | 5783 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
palmitoyl-CoA + mucus glycoprotein | Saccharomyces cerevisiae | protein S-palmitoylation is a posttranslational modification in which a palmitoyl group is added to a protein via a thioester linkage on cysteine | CoA + O-palmitoylglycoprotein | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
diverse strains | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged Erf2 with fused FLAG-tagged Erf4 from strain RJY1827, solubilization by 0.8% dodecylmaltoside and urea, and ultracentrifugation, purification by nickel affinity chromatography and desalting gel filtration, recombinant FLAG-tagged Erf2 by the same method | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
palmitoyl-CoA + mucus glycoprotein | protein S-palmitoylation is a posttranslational modification in which a palmitoyl group is added to a protein via a thioester linkage on cysteine | Saccharomyces cerevisiae | CoA + O-palmitoylglycoprotein | - |
r | |
palmitoyl-CoA + mucus glycoprotein | purified recombinant His6:Erf2-Erf4 complex | Saccharomyces cerevisiae | CoA + O-palmitoylglycoprotein | - |
r |
Synonyms | Comment | Organism |
---|---|---|
Pat | - |
Saccharomyces cerevisiae |
ras palmitoyl acyltransferase | - |
Saccharomyces cerevisiae |
Ras PAT | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
assay at | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
malfunction | in the absence of Erf4, the rate of hydrolysis of the active site palmitoyl thioester intermediate is increased, resulting in reduced palmitoyl transfer to a Ras2 substrate | Saccharomyces cerevisiae |
additional information | Erf4 and the C-terminal 58 amino acids of Erf2 are required for Erf2 stability. Role of the quality control system in degrading Erf2 molecules that are not in complex with Erf4. The C-terminal 58-amino acid residues of Erf2 are necessary and sufficient for ubiquitin-dependent, ERAD-mediated clearance of DHHC PATs | Saccharomyces cerevisiae |
physiological function | palmitoylation is a reversible modification involved in protein membrane targeting, receptor trafficking and signaling, vesicular biogenesis and trafficking, protein aggregation, and protein degradation. An example of the dynamic nature of this modification is the palmitoylation-depalmitoylation cycle that regulates the subcellular trafficking of Ras family GTPases. The Ras protein acyltransferase consists of a complex of Erf2-Erf4 in yeast cells. Both subunits Erf2 and Erf4 are required for PAT activity, one role of Erf4 is to regulate Erf2 stability through an ubiquitin-mediated pathway. Erf4 is required for the stable formation of the palmitoyl-Erf2 intermediate, the first step of palmitoyl transfer to protein substrates | Saccharomyces cerevisiae |