Recombinant Human GSTM4, GST-tagged

• Cat.No.: GSTM4-13577H
• Product Overview: Recombinant Human GSTM4 protein, fused to GST-tag, was expressed in E.coli and purified by GSH-sepharose.

Specification

• Species: Human
• Source: E.coli
• Tag: GST
• Protein Length: 1-218a.a.
• Description: This gene encodes a member of the GRB2/Sem5/Drk family and functions as a cytoplasmic signaling protein which contains an SH2 domain flanked by two SH3 domains. The SH2 domain interacts with ligand-activated receptors for stem cell factor and erythropoietin, and facilitates the formation of a stable complex with the BCR-ABL oncoprotein. This protein also associates with the Ras guanine nucleotide exchange factor SOS1 (son of sevenless homolog 1) through its N-terminal SH3 domain. In general, it couples signals from receptor and cytoplasmic tyrosine kinases to the Ras signaling pathway.
• Storage: The protein is stored in PBS buffer at -20℃. Avoid repeated freezing and thawing cycles.
• Storage Buffer: 1M PBS (58mM Na2HPO4,17mM NaH2PO4, 68mM NaCl, pH8. ) added with 100mM GSH and 1% Triton X-100,15%glycerol.

Gene Information

• Gene Name: GSTM4 glutathione S-transferase mu 4 [ Homo sapiens ]
• Official Symbol: GSTM4
• Synonyms: GSTM4; glutathione S-transferase mu 4; glutathione S transferase M4; glutathione S-transferase Mu 4; GST-Mu2; GTS-Mu2; GST class-mu 4; glutathione S-transferase M4; glutathione S-aryltransferase M4; glutathione S-alkyltransferase M4; glutathione S-aralkyltransferase M4; S-(hydroxyalkyl)glutathione lyase M4; GTM4; GSTM4-4; MGC9247; MGC131945
• Gene ID: 2948
• mRNA Refseq: NM_000850
• Protein Refseq: NP_000841
• MIM: 138333
• UniProt ID: Q03013
• Chromosome Location: 1p13
• Pathway: Biological oxidations, organism-specific biosystem; Drug metabolism - cytochrome P450, organism-specific biosystem; Drug metabolism - cytochrome P450, conserved biosystem; Glutathione conjugation, organism-specific biosystem; Glutathione metabolism, organism-specific biosystem; Glutathione metabolism, conserved biosystem; Metabolism, organism-specific biosystem
• Function: glutathione transferase activity; transferase activity

Citation

Biosynthetic metabolomes of cysteinyl-containing immunoresolvents

Journal: The FASEB Journal    PubMed ID: 31589826    Data: 2020/10/5

Authors: Charlotte C. Jouvene, Ashley E. Shay, Charles N. Serhan

Article Snippet:Recombinant human LTA 4 hydrolase (LTA 4 H), LTC 4 S, mGST2, and mGST3 were prepared as previously described in refs. 30 , 33 , and 34 , and TK05 (LTC 4 S inhibitor) was earlier identified through high-throughput screening ( 35 ).Recombinant human LTA 4 hydrolase (LTA 4 H), LTC 4 S, mGST2, and mGST3 were prepared as previously described in refs. 30 , 33 , and 34 , and TK05 (LTC 4 S inhibitor) was earlier identified through high-throughput screening ( 35 ).. Recombinant human GSTM4 was purchased from Creative BioMart (Shirley, NY, USA).. Synthetic eMaR, ePD, [ 13 C] 2 15 N-MCTR1, [ 13 C] 2 15 N-MCTR2, and [ 13 C] 2 15 N-MCTR3 were prepared by the lab of Dr. N. A. Petasis (University of Southern California, Los Angeles, CA, USA); their total organic synthesis will be reported separately.Synthetic eMaR, ePD, [ 13 C] 2 15 N-MCTR1, [ 13 C] 2 15 N-MCTR2, and [ 13 C] 2 15 N-MCTR3 were prepared by the lab of Dr. N. A. Petasis (University of Southern California, Los Angeles, CA, USA); their total organic synthesis will be reported separately.

Selective conversion of ePD and eMaR to PCTR1 and MCTR1 by specific human GST. Each human recombinant enzyme (LTC4S, mGST2, mGST3, and GSTM4; 1 μg/ml) was incubated with each of the 3 synthetic epoxides (25 μM) in 25 mM Tris-HCl (pH 8.0), 0.05% Triton X-100, and 5 mM reduced glutathione for 15 min at 37°C. LTC4, MCTR1, and PCTR1 were each identified and quantified by LC-MS/MS. A) Mechanism of the SN2 nucleophilic substitution reaction on the epoxide leading to the conversion to LTC4, PCTR1, and MCTR1 by these enzymes. B) Representative MRM chromatograms of LTC4, MCTR1, and PCTR1 produced by these enzymes. C) Percentage conversion of the epoxides by the 4 enzymes. Data are expressed as the percentages of each conjugate compound out of the total produced by 1 enzyme (n = 3). D) LTA4, eMaR, and ePD inhibit human LTA4H. LTA4H (100 ng/ml) was first incubated with LTA4 (5 μM), eMaR (5 μM), or ePD (5 μM) for 15 min and then with LTA4 (5 μM) for 15 min at 37°C for inhibition experiments. Top, quantification of LTB4 formation following the incubation of LTA4H and LTA4. Bottom, percentage inhibition of LTB4 formation by the 3 epoxides. Values are expressed as means ± sem, n = 3. *P < 0.05, **P < 0.01 (1-tailed Student’s t test vs. LTA4H + LTA4 condition).

Human M1- and M2-like macrophages express the enzymes involved in the biosynthesis of the CTRs. Human PBMCs were isolated from the whole blood of 3 individual donors and differentiated into M1-like and M2-like phenotypes. A) MFI of LTC4S, mGST2, mGST3, and GSTM4 in human PBMCs and M1- and M2-like macrophages. The gray dotted line represents the MFI for the isotype control. Values are represented as means ± sem. Two-way ANOVA with Tukey’s multiple comparisons test. M1- and M2-like compared with PBMC for each enzyme. *P < 0.05, ****P < 0.0001. B) Representative histograms of LTC4S, mGST2, mGST3, and GSTM4 expression in human M2-like macrophages as identified by flow cytometry at d 0 (undifferentiated PBMCs), d 4 and 6 (during differentiation), and d 8 (postpolarization).