The [alpha]-Ketoglutarate dehydrogenase complex (KGDHC) catalyzes the reaction of [alpha]- Ketoglutarate
with Coenzym A to succinyl-CoA under reduction of NAD to NADH.
Anti-mitochondrial antibodies (AMA) directed against the E2 subunit of pyruvate dehydrogenase (PDH-E2) which is a member of the inner mitochondrial membrane-expressed oxoacid dehydrogenase complex, and ketoglutarate
dehydrogenase are present in 95% of the patients3.
* Consists of pyridoxyl (50%) and alpha ketoglutarate
These patients have combined deficiency of BKCD, pyruvate and alpha ketoglutarate
This results in an increased conversion of glutamate to a ketoglutarate
and, thus, an increased flux of oxidative substrate into the mitochondria (anaplerosis).
The following methods were used: calcium: Arsenazo III dye method; uric acid: the uricase method; glucose: glucose oxidase method; aspartate aminotransferase (AST): aspartate plus the alpha ketoglutarate
method; and creatine kinase (CK): the creatine phosphate method.
Glutamate is acted upon by glutamate dehydrogenase generating alpha ketoglutarate
Also, glutamate dehydrogenase (EC 220.127.116.11) can be used as a further coupled-enzyme to form glutamic acid from ketoglutarate
and ammonia; the reaction requires NADH and can be monitored at 340 nm .
The final reaction volume of 1000 [micro]L contained 250 mM imidazole buffer, pH 6.5, 0.1 M KCl, 0.04 M Mg[Cl.sub.2], 6 mM [alpha]- ketoglutarate
, 20 [micro]L of glutamate dehydrogenase (1:5 aqua bidest), 0.15 mM NADPH, and 5, 10 or 20 [micro]L of muscle homogenate.
du Pont de Nemours and Company (Wilmington, DE) has patented four chimeric genes, a first encoding lysine-insensitive aspartokinase, which is operably linked to a plant chloroplast transit sequence, a second encoding lysine-insensitive dihydrodipicolinic acid synthase, which is operably linked to a plant chloroplast transit sequence, a third encoding a lysine-rich protein, and a fourth encoding a plant lysine ketoglutarate
reductase, all operably linked to plant seed-specific regulatory sequences.
"It contains our proprietary Matraxol Technology Arginine Alpha Ketoglutarate
, Cinnulin PF and LeucoSelect OPC," Hohn adds.