Nitrogen isotope signature evidences ammonium deprotonation as a common transport mechanism for the AMT-Mep-Rh protein superfamily

Science Advances
Idoia ArizCristina Cruz

Abstract

Ammonium is an important nitrogen (N) source for living organisms, a key metabolite for pH control, and a potent cytotoxic compound. Ammonium is transported by the widespread AMT-Mep-Rh membrane proteins, and despite their significance in physiological processes, the nature of substrate translocation (NH3/NH4+) by the distinct members of this family is still a matter of controversy. Using Saccharomyces cerevisiae cells expressing representative AMT-Mep-Rh ammonium carriers and taking advantage of the natural chemical-physical property of the N isotopic signature linked to NH4+/NH3 conversion, this study shows that only cells expressing AMT-Mep-Rh proteins were depleted in 15N relative to 14N when compared to the external ammonium source. We observed 15N depletion over a wide range of external pH, indicating its independence of NH3 formation in solution. On the basis of inhibitor studies, ammonium transport by nonspecific cation channels did not show isotope fractionation but competition with K+. We propose that kinetic N isotope fractionation is a common feature of AMT-Mep-Rh-type proteins, which favor 14N over 15N, owing to the dissociation of NH4+ into NH3 + H+ in the protein, leading to 15N depletion in the cell and allowing...Continue Reading

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Citations

Jan 29, 2020·The Journal of Biological Chemistry·Pascal GanzBenjamin Neuhäuser
Jul 15, 2020·ELife·Gordon WilliamsonArnaud Javelle
Aug 26, 2020·ELife·William J Allen, Ian Collinson
Nov 16, 2019·Scientific Reports·Pascal GanzBenjamin Neuhäuser
Nov 8, 2019·PLoS Pathogens·Bert van den BergJulian C Rutherford
May 1, 2021·Biomolecules·Marina VolkovaAlbert Guskov
Aug 17, 2021·Frontiers in Bioengineering and Biotechnology·Muhammad Junaid YousafIn-Jung Lee

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Methods Mentioned

BETA
fluorescence spectroscopy
electrophoresis

Software Mentioned

SPSS

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