Abstract
Glutamate dehydrogenase (GDH) plays an important role in the ammonium assimilation and nitrogen metabolism by catalyzing the reversible oxidative deamination of L-glutamate to α-ketoglutarate. In the present study, the potential functions of GDH in response to heat stress were explored in the scleractinian coral Pocillopora damicornis (designated as PdGDH). The cDNA of PdGDH contained an open reading frame of 1 611 bp encoding a polypeptide of 536 amino acids, which exhibited the highest sequence identity to GDH of Stylophora pistillata (96% identity), and the deduced PdGDH protein was predicted to contain one GdhA domain (from Val95 to Tyr525). The recombinant protein of PdGDH (rPdGDH) was expressed in Escherichia coli BL21 (DE3)-Transetta, and its catalytic activity was measured under different temperatures, pH conditions and epigallocatechin-3-gallate (EGCG, a GDH inhibitor) concentrations. The purified rPdGDH only used reduced coenzyme nicotinamide adenine dinucleotide (NADH) as coenzyme, and its highest activity was observed at 35 °C and pH 7.5, respectively. The rPdGDH activity was negatively correlated with the concentration of EGCG, and was inhibited by more than half (65%, P<0.05) at 10−4 mol/L EGCG. No significant alteration of PdGDH mRNA expression was detected at 12 h after exposure to heat and ammonium (P>0.05). Furthermore, the activities of NADH-GDH in the scleractinian coral P. damicornis increased significantly at 12 h after the heat and ammonium stress, and the NADH-GDH activity in the heat stress group (32.66 U/mg, P<0.05) was significantly higher than that in the heat and ammonium stress group (11.26 U/mg). These results collectively suggested that PdGDH, as a homologue of glutamate dehydrogenase in the scleractinian coral P. damicornis, could respond to heat stress at the protein level, which would have ability to further promote ammonium assimilation to increase the heat acclimatization of the coral-Symbiodiniaceae symbiotic association.
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Data Availability Statement
All data generated and/or analyzed during this study are available from the corresponding author upon request on reasonable request.
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The authors are grateful to all of the laboratory members for their continuous technical advice and helpful discussions.
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Supported by the Major Science and Technology Program of Hainan Province (No. ZDKJ2019011), the ISF-NSFC Joint Scientific Research Program (No. 42161144006 or 3511/21), the National Natural Science Foundation of China (Nos. 31772460, 42076145), and the Hainan Provincial Natural Science Foundation of China (Nos. 2019RC067, 420CXTD432)
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Tang, J., Cai, W., Yan, Z. et al. Involvement of the ammonium assimilation mediated by glutamate dehydrogenase in response to heat stress in the scleractinian coral Pocillopora damicornis. J. Ocean. Limnol. 40, 2001–2011 (2022). https://doi.org/10.1007/s00343-021-1266-8
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DOI: https://doi.org/10.1007/s00343-021-1266-8