Identification of the putative tumor suppressor Nit2 as omega-amidase, an enzyme metabolically linked to glutamine and asparagine transamination.
The present report identifies the enzymatic substrates of a member of the mammalian nitrilase-like (Nit) family. Nit2, which is widely distributed in nature, has been suggested to be a tumor suppressor protein. The protein was assumed to be an amidase based on sequence homology to other amidases and on the presence of a putative amidase-like active site. This assumption was recently confirmed by the publication of the crystal structure of mouse Nit2. However, the in vivo substrates were not previously identified. Here we report that rat liver Nit2 is omega-amidodicarboxylate amidohydrolase (E.C. 3.5.1.3; abbreviated omega-amidase), a ubiquitously expressed enzyme that catalyzes a variety of amidase, transamidase, esterase and transesterification reactions. The in vivo amidase substrates are alpha-ketoglutaramate and alpha-ketosuccinamate, generated by transamination of glutamine and asparagine, respectively. Glutamine transaminases serve to salvage a number of alpha-keto acids generated through non-specific transamination reactions (particularly those of the essential amino acids). Asparagine transamination appears to be useful in mitochondrial metabolism and in photorespiration. Glutamine transaminases play a particularly important role in transaminating alpha-keto-gamma-methiolbutyrate, a key component of the methionine salvage pathway. Some evidence suggests that excess alpha-ketoglutaramate may be neurotoxic. Moreover, alpha-ketosuccinamate is unstable and is readily converted to a number of hetero-aromatic compounds that may be toxic. Thus, an important role of omega-amidase is to remove potentially toxic intermediates by converting alpha-ketoglutaramate and alpha-ketosuccinamate to biologically useful alpha-ketoglutarate and oxaloacetate, respectively. Despite its importance in nitrogen and sulfur metabolism, the biochemical significance of omega-amidase has been largely overlooked. Our report may provide clues regarding the nature of the biological amidase substrate(s) of Nit1 (another member of the Nit family), which is a well-established tumor suppressor protein), and emphasizes a) the crucial role of Nit2 in nitrogen and sulfur metabolism, and b) the possible link of Nit2 to cancer biology.
Krasnikov
Boris F
BF
Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA. boris_krasnikov@nymc.edu
Chien
Chin-Hsiang
CH
Nostramo
Regina
R
Pinto
John T
JT
Nieves
Edward
E
Callaway
Myrasol
M
Sun
Jin
J
Huebner
Kay
K
Cooper
Arthur J L
AJ
D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000619:Aminohydrolases / Q000235:genetics; D000818:Animals; D001216:Asparagine / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005973:Glutamine / Q000378:metabolism*; D006801:Humans; D007656:Ketoglutaric Acids / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007656:Ketoglutaric Acids; D021122:Protein Subunits; D005973:Glutamine; C008230:alpha-ketoglutaramate; D001216:Asparagine; D000581:Amidohydrolases; C047679:omega-amidase; D000619:Aminohydrolases