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Encompassing Amino Acids: 17-669(end)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: The reaction mixture contained 100 mM Tris, pH 8.6, 150 mM K2HPO4, 25 mM L-glutamine, 2 mM NAD+, 4 units of GDH, and various amounts of GLS1 in a volume of 200 ?l. Reactions were initiated with the addition of GLS1, and the increase in NADH absorbance at 340 nm was continuously monitored for 7 minutes at 37°C. Molar extinction coefficient of NADH is 6,220 M-1cm-1.
Background: GLS1 is a phosphate-activated amidohydrolase that catalyzes the hydrolysis of glutamine to glutamate and ammonia. GLS1 is primarily expressed in the brain and kidney where it catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine. It plays an essential role in generating energy for metabolism, synthesizing the brain neurotransmitter glutamate and maintaining acid-base balance in the kidney. Alternate splicing results in multiple transcript variants.
Description: Human glutaminase (GLS), also known as glutaminase transcript variant 1 (GLS1), GAC, GAM and KGA, GenBank Accession No. NM_014905, a.a. 17-669(end), with N-terminal His-tag, MW 72.5 kDa, expressed in an E. coli cell expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20, and 20% glycerol.
Genbank: NM_014905
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag
Uniprot: O94925
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Van den Heuvel, A.P., et al., Cancer Biol. Ther. 2012;13(12):1185-1194.
2. Thangavelu, K., et al., Proc. Natl. Acad. Sci. USA 2012;109(2):7705-7710.
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