C12ORF5 Human, TAT

Chromosome 12 Open Reading Frame 5 Human-TAT
Cat. No.
BT13462
Source
Escherichia Coli.
Synonyms
Fructose-2,6-bisphosphatase TIGAR, TP53-induced glycolysis and apoptosis regulator, TIGAR, C12orf5.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis 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

TIGAR Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 283 amino acids (including the 270 residues of full-length TIGAR and a 13-residue C-terminal TAT peptide) and having a molecular mass of 31.7kDa. The TIGAR is purified by proprietary chromatographic techniques.

Product Specs

Introduction
C12ORF5, also known as TIGAR, is an enzyme induced by p53 that plays a crucial role in cellular metabolism and stress response. TIGAR acts by catalyzing the breakdown of fructose-2,6-bisphosphate (F-2,6-BP), a potent activator of glycolysis. By reducing F-2,6-BP levels, TIGAR shifts glucose metabolism away from glycolysis and towards the pentose phosphate pathway (PPP), leading to increased production of NADPH, an essential antioxidant molecule. This shift helps cells withstand oxidative stress and maintain redox balance. The protein transduction domain (PTD) of the HIV-1 TAT protein, known as TAT, facilitates the direct delivery of proteins into cells, enhancing their intracellular uptake.
Description
C12ORF5 Human Recombinant, fused with a TAT peptide for enhanced cellular uptake, is expressed in E. coli. This recombinant protein is a single, non-glycosylated polypeptide chain comprising 283 amino acids. The total amino acid count includes the 270 residues of full-length C12ORF5 and a 13-residue C-terminal TAT peptide. The molecular weight of the recombinant C12ORF5-TAT protein is 31.7 kDa. The purification process involves proprietary chromatographic techniques to ensure high purity.
Physical Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation
C12ORF5 was lyophilized from a 0.2µm filtered concentrated solution in 20mM PBS, pH 7.0, containing 350mM NaCl and 5% Trehalose.
Solubility
To reconstitute the lyophilized C12ORF5 protein, it is recommended to dissolve it in sterile 18 MΩ-cm H2O at a concentration of at least 100 µg/ml. The reconstituted solution can be further diluted in other aqueous solutions as needed.
Stability
Lyophilized C12ORF5 is stable at room temperature for up to 3 weeks when stored properly. It is crucial to store the lyophilized protein desiccated at a temperature below -18°C. After reconstitution, C12ORF5 should be stored at 4°C for a period of 2 to 7 days. For long-term storage, it is advised to add a carrier protein, such as 0.1% HSA or BSA, to the solution and store it at -18°C.
Repeated freeze-thaw cycles should be avoided to maintain protein stability.
Purity
The purity of C12ORF5 is determined using the following methods, which indicate a purity greater than 95.0%:
(a) Analysis by RP-HPLC (Reverse Phase High-Performance Liquid Chromatography).
(b) Analysis by SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis).
Biological Activity
The biological activity of recombinant human C12ORF5 is assessed based on its ability to shield U2OS cells from hydrogen peroxide-induced apoptosis. The specific activity is determined by pre-treating U2OS cells with varying concentrations of rHuC12ORF5-TAT (0.1-5.0 µg/ml) for 4 hours and then exposing them to hydrogen peroxide. The concentration range of rHuC12ORF5-TAT that confers protection against apoptosis is reported as its specific activity.
Synonyms
Fructose-2,6-bisphosphatase TIGAR, TP53-induced glycolysis and apoptosis regulator, TIGAR, C12orf5.
Source
Escherichia Coli.
Amino Acid Sequence
MARFALTVVR HGETRFNKEK IIQGQGVDEP LSETGFKQAA AAGIFLNNVK FTHAFSSDLM RTKQTMHGIL ERSKFCKDMT VKYDSRLRER KYGVVEGKAL SELRAMAKAA REECPVFTPP GGETLDQVKM RGIDFFEFLC QLILKEADQK EQFSQGSPSN CLETSLAEIF PLGKNHSSKV NSDSGIPGLA ASVLVVSHGA YMRSLFDYFL TDLKCSLPAT LSRSELMSVT PNTGMSLFII NFEEGREVKP TVQCICMNLQ DHLNGLTETR GGYGRKKRRQ RRR.

Product Science Overview

Discovery and Structure

The C12ORF5 gene was discovered in the early 2000s and has since been studied extensively. The protein encoded by this gene is known as TIGAR (TP53-Induced Glycolysis and Apoptosis Regulator). TIGAR is a newly discovered enzyme that largely regulates glucose breakdown in human cells . The protein has a tertiary structure similar to the histidine phosphatase fold and is approximately 30 kDa in size. The core of TIGAR is a β-sandwich composed of a six-stranded β-sheet encircled by four α-helices .

Function and Biological Role

TIGAR plays a crucial role in cellular metabolism by regulating the levels of fructose-2,6-bisphosphate, a key regulator of glycolysis. By lowering the levels of fructose-2,6-bisphosphate, TIGAR inhibits glycolysis and promotes the pentose phosphate pathway, which is essential for generating NADPH and ribose-5-phosphate . This shift in metabolic pathways helps cells to manage oxidative stress and repair DNA damage.

Additionally, TIGAR has the ability to protect cells from apoptosis (programmed cell death) induced by oxidative stress. This protective function is particularly important in cancer cells, where TIGAR activity can contribute to tumor survival and growth .

Implications in Cancer Research

The p53 tumor suppressor protein, which is activated in response to DNA damage, induces the expression of TIGAR. This connection between p53 and TIGAR has made the C12ORF5 gene a target of interest in cancer research. Studies have shown that TIGAR is overexpressed in various types of cancer, including breast, liver, and colorectal cancers . The overexpression of TIGAR in cancer cells helps them to survive under conditions of metabolic stress and contributes to their resistance to chemotherapy.

Researchers are exploring the potential of targeting TIGAR as a therapeutic strategy for cancer treatment. By inhibiting TIGAR activity, it may be possible to sensitize cancer cells to chemotherapy and reduce their ability to survive under metabolic stress .

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