Expression pattern and biochemical properties of zebrafish N-acetylglutamate synthase

PloS One
Ljubica CaldovicAlison Krufka

Abstract

The urea cycle converts ammonia, a waste product of protein catabolism, into urea. Because fish dispose ammonia directly into water, the role of the urea cycle in fish remains unknown. Six enzymes, N-acetylglutamate synthase (NAGS), carbamylphosphate synthetase III, ornithine transcarbamylase, argininosuccinate synthase, argininosuccinate lyase and arginase 1, and two membrane transporters, ornithine transporter and aralar, comprise the urea cycle. The genes for all six enzymes and both transporters are present in the zebrafish genome. NAGS (EC 2.3.1.1) catalyzes the formation of N-acetylglutamate from glutamate and acetyl coenzyme A and in zebrafish is partially inhibited by L-arginine. NAGS and other urea cycle genes are highly expressed during the first four days of zebrafish development. Sequence alignment of NAGS proteins from six fish species revealed three regions of sequence conservation: the mitochondrial targeting signal (MTS) at the N-terminus, followed by the variable and conserved segments. Removal of the MTS yields mature zebrafish NAGS (zfNAGS-M) while removal of the variable segment from zfNAGS-M results in conserved NAGS (zfNAGS-C). Both zfNAGS-M and zfNAGS-C are tetramers in the absence of L-arginine; addition...Continue Reading

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Citations

Dec 3, 2014·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Gabriel N AugheyJi-Long Liu
Jul 30, 2014·Molecular Genetics and Metabolism·B FeldmanL Caldovic
Feb 9, 2021·Frontiers in Physiology·Nantaporn HaskinsLjubica Caldovic

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Methods Mentioned

BETA
PCR

Software Mentioned

SYPRO Orange
GraphPad Prism
Thermofluor®
ORNT
MitoProt

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