Recombinant Full Length Human MGMT Protein, His tagged

• Cat.No.: MGMT-121H
• Product Overview: Recombinant Full Length Human MGMT Protein (1-207 aa) with His tag was expressed in E. coli.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Protein Length: 1-207 aa
• Description: Alkylating agents are potent carcinogens that can result in cell death, mutation and cancer. The protein encoded by this gene is a DNA repair protein that is involved in cellular defense against mutagenesis and toxicity from alkylating agents. The protein catalyzes transfer of methyl groups from O(6)-alkylguanine and other methylated moieties of the DNA to its own molecule, which repairs the toxic lesions. Methylation of the genes promoter has been associated with several cancer types, including colorectal cancer, lung cancer, lymphoma and glioblastoma.
• Molecular Mass: 23 kDa
• AASequence: MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEVPAPAAVLGGPEPLMQCTAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISYQQLAALAGNPKAARAVGGAMRGNPVPILIPCHRVVCSSGAVGNYSGGLAVKEWLLAHEGHRLGKPGLGGSSGLAGAWLKGAGATSGSPPAGRNHHHHHHHH
• Endotoxin: < 1 EU/μg by LAL
• Purity: > 90% by SDS-PAGE
• Storage: Store it under sterile conditions at -20 to -80 centigrade. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Storage Buffer: Sterile PBS, pH7.4
• Concentration: 0.5 mg/mL
• Publications:
  Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response (2016)
  Activatable Probes for Anti-Cancer Therapy (2019)

Gene Information

• Gene Name: MGMT O-6-methylguanine-DNA methyltransferase [ Homo sapiens (human) ]
• Official Symbol: MGMT
• Synonyms: MGMT; O-6-methylguanine-DNA methyltransferase; methylated-DNA--protein-cysteine methyltransferase; methylguanine-DNA methyltransferase; O-6-methylguanine-DNA-alkyltransferase; O6-methylguanine-DNA methyltransferase; 6-O-methylguanine-DNA methyltransferase
• Gene ID: 4255
• mRNA Refseq: NM_002412
• Protein Refseq: NP_002403
• MIM: 156569
• UniProt ID: P16455

Citation

Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

Journal: PLoS ONE    PubMed ID: 27035132    Data: 2016/4/1

Authors: Andrew A. Beharry, Zachary D. Nagel, Robert W Sobol

Article Snippet:Purified recombinant human MGMT (His-tagged, expressed in E . Coli ) was purchased from Creative BioMart.. All assays were carried out at 37°C in 70 mM HEPES buffer (pH 7.8) containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA.All assays were carried out at 37°C in 70 mM HEPES buffer (pH 7.8) containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA.

Fluorescence spectra showing the overall fold-changes in intensity observed when comparing fluorescence measured before (dashed line) and after (solid line) addition of purified MGMT protein are shown at left of each figure. Time courses (on the right) show time-dependent fluorescence increases immediately after addition of enzyme. Final probe and MGMT concentrations were 100 nM. Assays were run at 37°C in 70 mM HEPES buffer pH 7.8 containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA. (A) chemosensor 1 containing dT FAM , (B), chemosensor 2 containing Cy3, (C), chemosensor 3 containing dT TMR and (D), chemosensor 4 containing perylene nucleoside. Measurements were repeated 3 times. Standard deviations are provided in .

Incubation of purified MGMT enzyme with the inhibitors BG and PaTrin-2 led to a concentration dependent decrease in observed final fluorescence intensity, indicative of MGMT inhibition. MGMT (10 nM) was incubated with inhibitor for 10 min at 37°C in 70 mM HEPES buffer (pH 7.8) containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA. Final fluorescence was acquired 10 min after addition of probe (10 nM). Data were normalized to measurements without inhibitor. Each data point is the average of 3 measurements.