Product Description: Price

Glutamine fructose-6-phosphate amidotransferase (GFAT/GFPT) is the first enzyme involved in the ubiquitous hexosamine biosynthetic pathway (HBP) in eukaryotes and prokaryotes, catalyzing the conversion of glutamine and fructose-6-phosphate to glucosamine-6-phosphate and glutamate, which are mediated by the glutaminase domain and isomerase/transferase domain of GFAT, respectively. The HBP pathway is essential for aminosugar biosynthesis, leading to the formation of UDP-N-acetyl-glucosamine (UDP-GlcNAc), which participates in the production of proteoglycans, glycoproteins, polysaccharides, and glycolipids. It is estimated that between 1-3% of total cellular glucose is used for the synthesis of UDP-GlcNAc. There are two enzyme isoforms of GFAT (GFAT1 and GFAT2) encoded by two separate genes, gfpt1 and gfpt2, which share ~75% homology and differ in gene regulatory modes and tissue distribution. Disease relevance of GFAT is apparent from the findings that mutations in GFAT1 underlie congenital myasthenic syndrome characterized by a limb-girdle pattern of muscle weakness and that GFAT2 deficiency is linked to asthenozoospermia. Gfpt-specific SNPs are known risk factors of diabetes and increased GFAT activity is associated with carcinogenesis. Activity of eukaryotic GFAT is modulated through phosphorylation by PKA and AMPK. The assay kit takes advantage of the glutaminase activity of GFAT and relies on the detection of glutamate, which is coupled to the reduction of the tetrazolium salt INT in a NADH-coupled enzymatic reaction to formazan, which exhibits an absorption maximum at 492 nm.

#hexosamine #HBP #O-GlcNAc #OGA #OGT #O-GlcNAcylation

Kit Components:

GFAT Buffer: 3 ml, store at -80°C

GFAT Substrate: 1 ml, store at -80°C

Glutamine: 100 mg, store at 4ºC

TA Assay Solution: 5 ml (100 wells), store at -80ºC
10x Cell Lysis Solution: 25 ml, store at RT

SDS:

DMSO, INT, Tris, EDTA, GLDH

Related Kits:
Glucose Assay, GAPDH Assay, SORD Assay, Aldose Reductase Assay, GLS Assay