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Purification and expression of a processing protease on β‐alanine‐oxoglutarate aminotransferase from rat liver mitochondria
Author(s) -
Ohyama Tomoko,
Matsuda Koichi,
Tachibana Hideyuki,
Fujimoto Sakata Shigeko,
Mori Masataka,
Horiuchi Masahisa,
Tamaki Nanaya
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.07.043
Subject(s) - biochemistry , alanine , protein subunit , molecular mass , biology , peptide sequence , peptide , enzyme , microbiology and biotechnology , beta (programming language) , chemistry , amino acid , gene , computer science , programming language
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 β‐alanine‐oxoglutarate aminotransferase (β‐AlaAT I) in rats. Its molecular mass was 180 kDa as determined by gel filtration. A subunit molecular mass of 97 652 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.