Recombinant Full Length Saccharomyces Cerevisiae Palmitoyltransferase Akr1(Akr1) Protein, His-Tagged

• Cat.No.: RFL6209SF
• Product Overview: Recombinant Full Length Saccharomyces cerevisiae Palmitoyltransferase AKR1(AKR1) Protein (P39010) (1-764aa), fused to N-terminal His tag, was expressed in E. coli.

Specification

• Species: S.cerevisiae
• Source: E.coli
• Tag: His
• ProteinLength: Full Length (1-764)
• Form: Lyophilized powder
• AA Sequence: MVNELENVPRASTLTNEEQTVDPSNNDSQEDISLGDSNEITSLASLKAIRSGNEEESGNE QVNHNDEAEEDPLLTRYHTACQRGDLATVKEMIHGKLLEVNNDGDSTEHITGLHWASINN RLSVVDFLVSQGADVNARAGALHATPLHWAARYGYVYIVDFLLKHGADPTMTDDQGFNLL HLSVNSSNIMLVLYVLFNVVSKGLLDIDCRDPKGRTSLLWAAYQGDSLTVAELLKFGASI KIADTEGFTPLHWGTVKGQPHVLKYLIQDGADFFQKTDTGKDCFAIAQEMNTVYSLREAL THSGFDYHGYPIKKWFKKSQHAKLVTFITPFLFLGIAFALFSHINPLFVIIVLFLLAIAT NKGLNKFVLPSYGRMGVHNVTLLRSPLLSGVFFGTLLWVTIVWFFKVMPRTFSDEQYTNI LMLVILVSVFYLFGQLVIMDPGCLPEETDHENVRQTISNLLEIGKFDTKNFCIETWIRKP LRSKFSPLNNAVVARFDHYCPWIFNDVGLKNHKAFIFFITLMESGIFTFLALCLEYFDEL EDAHEDTSQKNGKCFILGASDLCSGLIYDRFVFLILLWALLQSIWVASLIFVQAFQICKG MTNTEFNVLMKESKSIGPDGLSFNENFNTTPEGFAPSIDPGEESNDTVLAPVPGSTIRKP RTCFGVCYAVTGMDQWLAVIKETIGIKDSTGHNVYSITSRIPTNYGWKRNVKDFWLTSDI NAPLWRRILYPPSGSKALLNGIEVDYFKLYKLPNKDVEQGNDMV
• Purity: Greater than 90% as determined by SDS-PAGE.
• Applications: SDS-PAGE
• Notes: Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
• Storage: Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
• Storage Buffer: Tris/PBS-based buffer, 6% Trehalose, pH 8.0
• Reconstitution: We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20℃/-80℃. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Gene Information

• Gene Name: AKR1
• Synonyms: AKR1; YDR264C; D9954.9; YD9230B.03C; Palmitoyltransferase AKR1; Ankyrin repeat-containing protein AKR1
• UniProt ID: P39010

Case Study

Case 1: Zhang ZT, et al. Biochim Biophys Acta Gen Subj. 2017

This study explores how Akr1, a palmitoyltransferase, mitigates methylmercury toxicity in yeast by identifying Meh1 as a key substrate that reduces this toxicity. Researchers utilized gene disruption and mutagenesis to link methylmercury's effects with vacuolar function and confirmed palmitoylation using the acyl-biotinyl exchange method. Results indicated that Meh1, when palmitoylated by Akr1, is essential for detoxification as part of the EGO complex, which inhibits autophagy. Methylmercury-induced vacuole deformation was more pronounced in yeast lacking EGO complex components but was lessened by the autophagy inhibitor 3-methyladenine, which also restored methylmercury sensitivity in Meh1-deficient yeast.

Fig1. Effect of deleting the Akr1-binding protein.

Fig2. Effect of mutation of Meh1 on the localization of Meh1.

Case 2: Politis EG, et al. J Biol Chem. 2005

The DHHC protein family, known for its DHHC cysteine-rich domain, includes potential palmitoyl acyl transferases (PATs) like Akr1p and Erf2p/Erf4p. This study determined the transmembrane topology of the yeast PAT Akr1p to identify cytoplasm-oriented domains involved in palmitoylation. Researchers inserted parts of the invertase protein Suc2p at various sites within Akr1p. Glycosylation of three resulting fusion proteins indicated luminal orientation, while the lack of glycosylation in seven others suggested a cytoplasmic location. This supports a model where Akr1p spans the membrane six times, with hydrophilic domains, including the ankyrin repeat-containing N-terminal and the DHHC-CRD, facing the cytoplasm. Only insertions at four cytoplasmic sites affected Akr1p function, highlighting the importance of these domains in its PAT activity.

Fig1. Analysis of N-linked glycosylation for Akr1p and Ste2p.

Fig2. PAT function of AKR1-SUC2-AKR1 alleles.

Application

Palmitoyltransferase is an important enzyme that plays a variety of roles in cell biology. This enzyme is responsible for catalyzing the palmitoylation of proteins, which is the addition of palmitoyl groups to the cysteine residues of proteins to form thioester bonds. This process is essential for protein localization, stability and function. The application of recombinant full-length Saccharomyces cerevisiae Palmitoyltransferase in practical life mainly includes:

Biopharmaceutical: In the production of recombinant proteins, this enzyme can be used to improve protein stability and function, especially in the production of therapeutic proteins and vaccines.

Metabolic engineering: Metabolic engineering of saccharomyces cerevisiae can enhance its ability to produce specific compounds, such as beta-carotene, by regulating the fatty acid synthesis pathway to increase production.

Food industry: In the food industry, this enzyme may help to improve the nutritional properties of foods, for example by altering fatty acid composition to produce healthier food ingredients.

Cosmetics industry: In cosmetics, palmitoyl transferase may be used to produce specific types of lipids that improve skin health and the performance of products.

Research tool: In basic biology research, this enzyme can be used as a tool to study protein palmitoylation, helping scientists better understand cell signaling and protein function.

Drug screening: In drug development, palmitoyl transferase can be used as a drug target to screen for small molecule compounds that can regulate their activity.

Fig1. Protein palmitoylation dynamically regulates protein function and signal transduction. (Xiaoyuan Yang, 2020)