GPT Rat

Glutamic-Pyruvate Transaminase Rat Recombinant
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Cat. No.
BT1522
Source
E.coli.
Synonyms
Alanine aminotransferase 1 (EC:2.6.1.2), ALT1, Glutamate pyruvate transaminase 1, GPT 1, Glutamic--alanine transaminase 1, Glutamic--pyruvic transaminase 1, Gpt, Aat1, Gpt1.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 90.0% 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

Biological Functions

  • Essential for gluconeogenesis and amino acid metabolism

  • Serum biomarker for hepatocellular damage

  • Expressed in liver (highest), kidney, heart, and skeletal muscle

Strain Development

The gpt delta rat system, first developed by Nohmi et al., contains lambda EG10 phage DNA (5-10 copies) integrated into chromosome 4 (F344 strain) or chromosome 1 (Wistar Hannover strain) . Key advancements:

GenerationBackground StrainCopy NumberPackaging Efficiency*Homozygosity
Original F3445-101×10^5 pfu/reactionHeterozygous
WH-Homozygous Wistar Hannover≥308×10^5 pfu/reactionHomozygous

*Measured via lambda phage packaging from liver genomic DNA

Mutation Detection Capabilities

The model enables simultaneous analysis of:

  • gpt assay: 6-thioguanine selection for point mutations

  • Spi- assay: Deletion detection (>1kb) through gam/red gene inactivation

Experimental validation with mutagens:

TreatmentDoseLiver gpt MF (×10^-6)Bone Marrow gpt MF (×10^-6)Spi- Frequency (×10^-6)
Control -3.2 ± 0.52.8 ± 0.30.9 ± 0.2
ENU (200 mg/kg)Single IP18.4 ± 2.1*15.6 ± 1.8*1.1 ± 0.3
BaP (30 mg/kg)5× oral gavage12.7 ± 1.6*9.4 ± 1.1*3.8 ± 0.7*

*Significant increase vs control (p<0.01)

Toxicological Studies

  • Detected liver GST-P+ foci formation in F344 gpt delta rats treated with hepatocarcinogens (aflatoxin B1, 2-AAF)

  • Identified dose-dependent suppression of colon tumors by silymarin (milk thistle extract) in DMH/DSS-induced carcinogenesis

Genotoxicity Testing

Advantages over conventional models:

  • Simultaneous assessment of general toxicity and mutagenicity

  • 89% concordance with carcinogenicity data in 56 chemical evaluations

  • Enables organ-specific mutation spectrum analysis

Limitations

  • Background mutation frequency varies by tissue (liver: 3.2×10^-6, kidney: 2.1×10^-6)

  • Requires 100-200 mg tissue samples for reliable phage packaging

  • Lower transgene recovery vs murine models (40% efficiency compared to mice)

Optimization Strategies

  • Use of homozygous WH strain increases DNA yield 8-fold

  • Carrier proteins (0.1% HSA) enhance recombinant GPT stability during freeze-thaw

  • Triple packaging reactions recommended for low-copy F344 strains

Product Specs

Introduction
GPT (Glutamate Pyruvate Transaminase) plays a critical role in the metabolism of glucose and amino acids by catalyzing the reversible conversion of alanine and 2-oxoglutarate to pyruvate and glutamate. Its activity is widely used as a clinical marker for liver health and potential damage.
Description
Recombinant GPT from Rat, expressed in E. coli, is a single, non-glycosylated polypeptide chain. It consists of 519 amino acids, with the first 496 representing the GPT protein, and has a molecular weight of 57.5 kDa. The protein includes a 23 amino acid His-tag at the N-terminus and is purified using proprietary chromatographic methods.
Physical Appearance
A clear, colorless solution that has been sterilized by filtration.
Formulation
The GPT protein is provided in a solution at a concentration of 1 mg/ml. The solution is formulated with Phosphate Buffered Saline at a pH of 7.4 and contains 10% glycerol.
Stability
For short-term storage (up to 4 weeks), the product can be stored at 4°C. For longer storage, it's recommended to freeze the product at -20°C. Adding a carrier protein such as HSA or BSA (0.1%) is advised for extended storage. Avoid repeated freezing and thawing.
Purity
The purity of the GPT protein is greater than 90% as determined by SDS-PAGE analysis.
Biological Activity
The specific activity is measured as greater than 60 units/mg. This is defined as the amount of enzyme required to catalyze the conversion of 1 micromole of L-Alanine to L-Glutamate per minute at a pH of 7.5 and a temperature of 37°C.
Synonyms
Alanine aminotransferase 1 (EC:2.6.1.2), ALT1, Glutamate pyruvate transaminase 1, GPT 1, Glutamic--alanine transaminase 1, Glutamic--pyruvic transaminase 1, Gpt, Aat1, Gpt1.
Source
E.coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSMASRVND QSQASRNGLK GKVLTLDTMN PCVRRVEYAV RGPIVQRALE LEQELRQGVK KPFTEVIRAN IGDAQAMGQR PITFFRQVLA LCVYPNLLSS PDFPEDAKRR AERILQACGG HSLGAYSISS GIQPIREDVA QYIERRDGGI PADPNNIFLS TGASDAIVTM LKLLVSGEGR ARTGVLIPIP QYPLYSAALA ELDAVQVDYY LDEERAWALD IAELRRALCQ ARDRCCPRVL CVINPGNPTG QVQTRECIEA VIRFAFKEGL FLMADEVYQD NVYAEGSQFH SFKKVLMEMG PPYSTQQELA SFHSVSKGYM GECGFRGGYV EVVNMDAEVQ KQMGKLMSVR LCPPVPGQAL MDMVVSPPTP SEPSFKQFQA ERQEVLAELA AKAKLTEQVF NEAPGIRCNP VQGAMYSFPQ VQLPLKAVQR AQELGLAPDM FFCLCLLEET GICVVPGSGF GQQEGTYHFR MTILPPMEKL RLLLEKLSHF HAKFTHEYS.

Product Science Overview

Structure and Source

The recombinant form of GPT from rats is a bioactive protein with a molecular weight of approximately 55 kDa . It is typically produced in E. coli and includes a His-tag for purification purposes . The enzyme consists of 496 amino acids and is found in various tissues, including the liver, kidney, heart, and skeletal muscles .

Function and Mechanism

GPT’s primary function is to facilitate the reversible conversion of L-alanine and alpha-ketoglutarate to L-glutamate and pyruvate . This reaction is essential for the metabolism of nitrogen and the production of glucose in the liver. The enzyme’s activity is crucial for maintaining the balance of amino acids and energy production in cells .

Clinical Significance

GPT is widely used as a biomarker for liver function. Elevated levels of GPT in the blood can indicate liver damage or disease, such as hepatitis, cirrhosis, or liver cancer . It is also used in research to study various diseases and disorders, including liver disease, glucose intolerance, and viral hepatitis .

Applications

Recombinant GPT is used in various research applications, including enzyme activity assays, protein-protein interaction studies, and drug development . Its high purity and specific activity make it a valuable tool for studying the biochemical pathways involving amino acid metabolism and liver function .

Storage and Stability

Recombinant GPT should be stored at 4°C for short-term use and at -20°C for long-term storage to maintain its stability and activity . It is typically supplied in a buffer containing PBS (pH 7.4) and 10% glycerol to prevent degradation .

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