MGMT Human

O-6-Methylguanine-DNA Methyltransferase Human Recombinant
Cat. No.
BT10458
Source
Escherichia Coli.
Synonyms
Methylated-DNA--protein-cysteine methyltransferase, 6-O-methylguanine-DNA methyltransferase, O-6-methylguanine-DNA-alkyltransferase, MGMT.
Appearance
Sterile Filtered clear colorless solution.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

MGMT Human Recombinant fused to a 20 amino acid His Tag at N-terminal produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 227 amino acids (1-207) and having a molecular mass of 23.8 kDa.
The MGMT is purified by proprietary chromatographic techniques.

Product Specs

Introduction
MGMT is an enzyme crucial for repairing O-6-methylguanine, a DNA base prone to damage by alkylating agents. This repair process is vital for cellular defense against the harmful effects of O-6-methylguanine. MGMT functions by transferring the alkyl group from the damaged guanine to a cysteine residue within itself. Notably, variations in MGMT expression levels are linked to the prognosis of solid cancers in humans, with decreased expression often correlating with methylation. Human MGMT acts as a negative regulator of estrogen receptor-mediated transcription in response to alkylation-induced DNA damage. Furthermore, hypermethylation of the MGMT promoter is implicated in the early stages of colorectal carcinogenesis and is also associated with oral squamous cell carcinomas.
Description
This product consists of recombinant human MGMT, with an N-terminal 20 amino acid His Tag, produced in E. coli. It exists as a single, non-glycosylated polypeptide chain comprising 227 amino acids (residues 1-207) and possesses a molecular weight of 23.8 kDa. The purification of MGMT is achieved using proprietary chromatographic techniques.
Physical Appearance
The product appears as a clear, colorless solution that has been sterilized by filtration.
Formulation
The MGMT solution is prepared in a buffer consisting of 20mM Tris-HCl at pH 7.5, 1mM DTT, and 10% glycerol.
Stability
For short-term storage (2-4 weeks), the product should be kept at 4°C. For extended storage, freezing at -20°C is recommended. The addition of a carrier protein (0.1% HSA or BSA) is advisable for long-term storage. Repeated freezing and thawing cycles should be avoided.
Purity
The purity of this product exceeds 95.0%, as determined by SDS-PAGE analysis.
Synonyms
Methylated-DNA--protein-cysteine methyltransferase, 6-O-methylguanine-DNA methyltransferase, O-6-methylguanine-DNA-alkyltransferase, MGMT.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MDKDCEMKRT TLDSPLGKLE LSGCEQGLHE IKLLGKGTSA ADAVEVPAPA AVLGGPEPLM QCTAWLNAYF HQPEAIEEFP VPAFHHPVFQ QESFTRQVLW KLLKVVKFGE VISYQQLAAL AGNPKAARAV GGAMRGNPVP ILIPCHRVVC SSGAVGNYSG GLAVKEWLLA HEGHRLGKPG LGGSSGLAGA WLKGAGATSG SPPAGRN.

Product Science Overview

Introduction

O-6-Methylguanine-DNA Methyltransferase (MGMT), also known as methylguanine methyltransferase, is a crucial enzyme involved in the DNA repair mechanism. This enzyme specifically repairs the O-6 position of guanine, which can be methylated due to exposure to alkylating agents. The human recombinant form of MGMT is produced through recombinant DNA technology, allowing for its use in various research and therapeutic applications.

Biological Significance

MGMT plays a pivotal role in maintaining genomic stability by removing alkyl groups from the O-6 position of guanine, thereby preventing mutations that could lead to cancer . The enzyme’s activity is essential for protecting cells from the mutagenic, carcinogenic, and cytotoxic effects of DNA alkylation . In cancer therapy, MGMT’s role is particularly significant as it can influence the effectiveness of alkylating chemotherapeutic agents .

Mechanism of Action

MGMT repairs DNA by transferring the alkyl group from the O-6 position of guanine to a cysteine residue within its own structure. This reaction is a one-time event for each MGMT molecule, as the enzyme is irreversibly inactivated after the transfer. This unique mechanism underscores the importance of MGMT in cellular defense against DNA damage .

Clinical Relevance

The activity of MGMT is a critical factor in the response to alkylating agents used in chemotherapy, such as temozolomide and dacarbazine . Tumors with high MGMT activity can repair the DNA damage caused by these agents, leading to resistance. Conversely, tumors with low MGMT activity are more susceptible to these treatments. Therefore, assessing MGMT activity or promoter methylation status can be a valuable prognostic marker in cancer therapy .

Research and Therapeutic Applications

Human recombinant MGMT is utilized in research to study DNA repair mechanisms and to develop strategies to modulate its activity for therapeutic purposes. For instance, inhibiting MGMT activity can enhance the efficacy of alkylating agents in cancer treatment. Additionally, understanding the regulation of MGMT expression and activity can provide insights into overcoming drug resistance in cancer therapy .

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