Recombinant Escherichia coli 8-oxo-dGTP diphosphatase (mutT)

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Code CSB-EP357355ENV
Size US$388
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Product Details

Purity
Greater than 90% as determined by SDS-PAGE.
Target Names
mutT
Uniprot No.
Research Area
Others
Alternative Names
mutT; b0099; JW0097; 8-oxo-dGTP diphosphatase; 8-oxo-dGTPase; EC 3.6.1.55; 7,8-dihydro-8-oxoguanine-triphosphatase; Mutator protein MutT; dGTP pyrophosphohydrolase
Species
Escherichia coli (strain K12)
Source
E.coli
Expression Region
1-129aa
Target Protein Sequence
MKKLQIAVGIIRNENNEIFITRRAADAHMANKLEFPGGKIEMGETPEQAVVRELQEEVGITPQHFSLFEKLEYEFPDRHITLWFWLVERWEGEPWGKEGQPGEWMSLVGLNADDFPPANEPVIAKLKRL
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
30.9kDa
Protein Length
Full Length
Tag Info
N-terminal 6xHis-SUMO-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is 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°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

The Recombinant E.coli K12 mutT protein is a protein encoded by recombinant DNA that was cloned in an expression vector that supported the expression of mutT gene. This recombinant mutT protein was expressed in the host. The expression region is 1-129aa of the E.coli K12 mutT. In the production, the expression vector contains N-terminal 6xHis-SUMO tag. Every production step was performed with a strict QC system. The purity of this protein is 90%+ determined by SDS-PAGE.

The MutT protein in Escherichia coli removes 8-oxo-deoxyguanosine triphosphate (8-oxo-dGTP) and 8-oxo-guanosine triphosphate (8-oxo-GTP) from the nucleotide pools precluding incorporation into DNA and RNA. 8-oxo-dGTP is a potent mutagenic nucleotide that is readily incorporated into DNA opposite template C or A, with a preference for template A. Therefore, by cleaning the 8-oxo-dGTP pool, MutT reduces spontaneous transversion mutations ∼1000-fold. In addition, MutT can also act on 8-oxo-guanosine triphosphate (8-oxo-GTP) and convert it to 8-oxo-GMP averting mutagenic nucleotide incorporation into RNA. mutT mutants are known to exhibit phenotypes independent of DNA mutational effects, which have been attributed to a decrease in transcription fidelity. The persistence of oxidized ribonucleotides in the available nucleotide pool and the subsequent incorporation of 8-oxo-GTP into mRNA, causing T to G transversions in the nascent transcript, was assumed to be the mechanism for RNA errors and these phenotypes. It was suggested that the absence of MutT can increase the readthrough of a stop codon mutation through 8-oxo-GTP incorporation generating a 30-fold increase in functional protein levels in mutT cultures compared to wild-type cultures. Such RNA infidelity in mutT strains may account for the accumulation of misfolded proteins and the observed cytotoxicity of aminoglycoside antibiotics. mutT phenotypes, such as protein mistranslation or antibiotic sensitivity, that have been attributed to 8-oxo-GTP require other explanations than simply 8-oxo-GTP misincorporation into mRNA.

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Target Background

Function
Specifically hydrolyzes both 8-oxo-deoxyguanosine triphosphate (8-oxo-dGTP) and 8-oxo-guanosine triphosphate (8-oxo-GTP) to the related monophosphates, thereby cleaning up the nucleotide pools and preventing misincorporation of 8-oxoGua into DNA and RNA. It prevents replicational errors by removing an oxidatively damaged form of guanine (8-oxo-dGTP) from DNA and the nucleotide pool. 8-oxo-dGTP can be inserted opposite dA and dC residues of template DNA with almost equal efficiency thus leading to A.T to G.C transversions. MutT may also ensure transcriptional fidelity, removing 8-oxo-GTP from the ribonucleotide triphosphate pool. However, due to the lower efficiency of RNA polymerase 8-oxo-GTP incorporation, MutT is probably not a major contributor to transcriptional fidelity. It also hydrolyzes 8-oxo-dGDP and 8-oxo-GDP to their monophosphate form. In vitro, can also use dGTP, dGDP and other various nucleoside di- and triphosphates, with much lower efficiency. Works cooperatively with MutM and MutY to prevent accumulation in the DNA of oxidized guanine residues.
Gene References into Functions
  1. The study investigated the role of MutT on transcription fidelity and find no increase in epigenetic switch frequency in the absence of MutT function. PMID: 25294823
  2. MutT mutation increased the point-mutation rate 150-fold. PMID: 23248287
  3. The MutT functions are not needed under anaerobic condition, yet the level of the MutT protein in cell is kept constant, probably for preparing for sudden changes of oxygen pressure. PMID: 21147134
  4. MutT hydrolyzes four types of 8-oxoGua (8-oxo-7,8-dihydroguanine)-containing nucleoside diphosphates and triphosphates to the related monophosphates, thereby preventing misincorporation of 8-oxoGua into DNA and RNA. PMID: 15850400
  5. transient state kinetic analysis of the reaction pathway of mutT; findings confirm an iso kinetic mechanism and explain the preferred hydrolysis of 8-oxo-dGTP PMID: 16285737
  6. These results indicate that mutT plays important roles in mutagenesis induced by 2-OH-dATP and 8-OH-dGTP in vivo. PMID: 17150583
  7. The mutT defect does not elevate chromosomal fragmentation in Escherichia coli because of the surprisingly low levels of MutM/MutY-recognized DNA modifications. PMID: 17616589

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Protein Families
Nudix hydrolase family
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