Purification and expression of a processing protease on beta-alanine-oxoglutarate aminotransferase from rat liver mitochondria.
GABA[arrow beta]AlaAT convertase is an endopeptidase that processes brain-type 4-aminobutyrate aminotransferase (GABA AT; EC 2.6.1.19) to liver-type beta-alanine-oxoglutarate aminotransferase (beta-AlaAT I) in rats. Its molecular mass was 180 kDa as determined by gel filtration. A subunit molecular mass of 97652 Da was measured using MALDI-TOF MS. The N-terminal sequence of the purified GABA[arrow beta]AlaAT convertase was SRVEVSKVLILGSGGLSIGQAGEFDYSGSQAV- and was identical to residues 418-449 of carbamoyl-phosphate synthetase I (CPS I; EC 1.2.1.27) purified from rat liver. The subunit molecular mass and the N-terminal amino acid sequence suggested that GABA[arrow beta]AlaAT convertase was the 418-1305 peptide of CPS I. An expression vector containing the coding region of the 418-1305 peptide of rat CPS I was transfected into NIH3T3 cells and the extract of the cells showed GABA[arrow beta]AlaAT convertase activity.
Faculty of Nutrition and High Technology Research Center, Kobe-Gakuin University, Arise 518, Ikawadani-cho, Nishi-ku, Kobe 651-2180, Japan. f9blh801@nutr.kobegakuin.ac.jp
Tomoko
T
Ohyama
Koichi
K
Matsuda
Hideyuki
H
Tachibana
Shigeko
S
Fujimoto Sakata
Masataka
M
Mori
Masahisa
M
Horiuchi
Nanaya
N
Tamaki
D016475:3T3 Cells; D000410:Alanine Transaminase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008930:Mitochondria, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013552:Swine; D000637:Transaminases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D011994:Recombinant Proteins; D000637:Transaminases; C490713:beta-alanine-oxoglutarate aminotransferase, rat; D000410:Alanine Transaminase; D010450:Endopeptidases