GOT2, also known as glutamate oxaloacetate transaminase 2, belongs to the class-I pyridoxal- phosphate-dependent aminotransferase family. glutamate oxaloacetate transaminas is a pyridoxal phosphate- dependent enzyme which exists in cytoplasmic and inner-membrane mitochondrial forms, GOT1 and GOT2, respectively. GOT plays a role in amino acid metabolism and the urea and tricarboxylic acid cycles. The two enzymes are homodimeric and show close homology. Source: Recombinant protein corresponding to aa30-430 from human GOT2, fused to His-tag at N-terminal expressed in E. coli. Molecular Weight: ~47kD (424aa) Specific Activity: >60units/mg, and is defined as the amount of enzyme that convert 1umole of a-ketoglutarate to L-Glutamate/minute at pH 8.0 at 25C. Activity Assay: 1. Prepare a 180ul assay buffer into a suitable container. The concentrations are 110mM Tris-HCl (pH8.0), 0.44mM beta-nicotinamide adenine dinucleotide, reduced form, 55mM L-aspartate, 2.75mM alpha-ketoglutaric acid, 82.5 units MDH. 2. Equilibrate to 25C and monitor at A340nm until the value is constant using a spectrophotometer. 3. Add 20ul of recombinant GOT2 protein 2ug/ml in assay buffer. Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MGSSSWWTHV EMGPPDPILG VTEAFKRDTN SKKMNLGVGA YRDDNGKPYV LPSVRKAEAQ IAAKNLDKEY LPIGGLAEFC KASAELALGE NSEVLKSGRF VTVQTISGTG ALRIGASFLQ RFFKFSRDVF LPKPTWGNHT PIFRDAGMQL QGYRYYDPKT CGFDFTGAVE DISKIPEQSV LLLHACAHNP TGVDPRPEQW KEIATVVKKR NLFAFFDMAY QGFASGDGDK DAWAVRHFIE QGINVCLCQS YAKNMGLYGE RVGAFTMVCK DADEAKRVES QLKILIRPMY SNPPLNGARI AAAILNTPDL RKQWLQEVKV MADRIIGMRT QLVSNLKKEG STHNWQHITD QIGMFCFTGL KPEQVERLIK EFSIYMTKDG RISVAGVTSS NVGYLAHAIH QVTK Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
