Biochemical properties of recombinant human and mouse N-acetylglutamate synthase.
N-Acetylglutamate synthase (NAGS, EC 2.3.1.1) is a mitochondrial enzyme that catalyzes the formation of N-acetylglutamate (NAG) from glutamate and acetylcoenzyme A. NAG is an obligatory activator of carbamylphosphate I (CPSI), the first and a rate limiting enzyme of ureagenesis. The enzymatic activity of NAGS increases in the presence of arginine. Since the level of NAGS activity depends on the concentrations of two amino acids, glutamate and arginine, and it supplies the essential cofactor for CPSI, NAGS may play an important role in the regulation of ureagenesis. The amino acid sequences of human and mouse NAGS consist of three regions with different degrees of conservation: the mitochondrial targeting signal (MTS), the variable domain, and the conserved domain. Removal of the MTS results in mature NAGS (NAGS-M) while removal of the MTS and the variable domain results in conserved NAGS (NAGS-C). The biochemical properties of purified recombinant human and mouse NAGS-M and NAGS-C were determined in this study with the goal of better understanding the role of the variable domain in NAGS function. The activity of all four proteins doubled in the presence of arginine, while the affinities for substrates changed less than two fold. The turnover numbers of NAGS-C are double those of NAGS-M proteins. Processing of NAGS-M to form NAGS-C results in an enzyme with higher catalytic activity and could play a role in the regulation of NAG production, CPSI function, and urea synthesis.
Caldovic
Ljubica
L
Children's Research Institute, Children's National Medical Center, The George Washington University, Washington, DC 20010, USA.
Lopez
Giselle Y
GY
Haskins
Nantaporn
N
Panglao
Maria
M
Shi
Dashuang
D
Morizono
Hiroki
H
Tuchman
Mendel
M
D051046:Amino-Acid N-Acetyltransferase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D051379:Mice; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D001120:Arginine; D051046:Amino-Acid N-Acetyltransferase; C494874:NAGS protein, human; C494877:Nags protein, mouse