A conserved cysteine near the P-site is accessible to cysteine modifications and increases ROS stability in P-type plasma membrane H+-ATPase
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A conserved cysteine near the P-site is accessible to cysteine modifications and increases ROS stability in P-type plasma membrane H+-ATPase. / Welle, Marcel; Pedersen, Jesper Torbøl; Ravnsborg, Tina; Hayashi, Maki; Maaß, Sandra; Becher, Dörte; Jensen, Ole Nørregaard; Stöhr, Christine; Palmgren, Michael.
I: Biochemical Journal, Bind 478, Nr. 3, 2021, s. 619-632.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - A conserved cysteine near the P-site is accessible to cysteine modifications and increases ROS stability in P-type plasma membrane H+-ATPase
AU - Welle, Marcel
AU - Pedersen, Jesper Torbøl
AU - Ravnsborg, Tina
AU - Hayashi, Maki
AU - Maaß, Sandra
AU - Becher, Dörte
AU - Jensen, Ole Nørregaard
AU - Stöhr, Christine
AU - Palmgren, Michael
N1 - Copyright 2021 The Author(s).
PY - 2021
Y1 - 2021
N2 - Sulfur-containing amino acid residues function in antioxidative responses, which can be induced by the reactive oxygen species generated by excessive copper and hydrogen peroxide. In all Na+/K+, Ca2+, and H+ pumping P-type ATPases, a cysteine residue is present two residues upstream of the essential aspartate residue, which is obligatorily phosphorylated in each catalytic cycle. Despite its conservation, the function of this cysteine residue was hitherto unknown. In this study, we analyzed the function of the corresponding cysteine residue (Cys-327) in the autoinhibited plasma membrane H+-ATPase isoform 2 (AHA2) from Arabidopsis thaliana by mutagenesis and heterologous expression in a yeast host. Enzyme kinetics of alanine, serine, and leucine substitutions were identical to those of the wild-type pump but the sensitivity of the mutant pumps was increased towards copper and hydrogen peroxide. Peptide identification and sequencing by mass spectrometry demonstrated that Cys-327 was prone to oxidation. These data suggest that Cys-327 functions as a protective residue in the plasma membrane H+-ATPase, and possibly in other P-type ATPases as well.
AB - Sulfur-containing amino acid residues function in antioxidative responses, which can be induced by the reactive oxygen species generated by excessive copper and hydrogen peroxide. In all Na+/K+, Ca2+, and H+ pumping P-type ATPases, a cysteine residue is present two residues upstream of the essential aspartate residue, which is obligatorily phosphorylated in each catalytic cycle. Despite its conservation, the function of this cysteine residue was hitherto unknown. In this study, we analyzed the function of the corresponding cysteine residue (Cys-327) in the autoinhibited plasma membrane H+-ATPase isoform 2 (AHA2) from Arabidopsis thaliana by mutagenesis and heterologous expression in a yeast host. Enzyme kinetics of alanine, serine, and leucine substitutions were identical to those of the wild-type pump but the sensitivity of the mutant pumps was increased towards copper and hydrogen peroxide. Peptide identification and sequencing by mass spectrometry demonstrated that Cys-327 was prone to oxidation. These data suggest that Cys-327 functions as a protective residue in the plasma membrane H+-ATPase, and possibly in other P-type ATPases as well.
U2 - 10.1042/BCJ20200559
DO - 10.1042/BCJ20200559
M3 - Journal article
C2 - 33427868
VL - 478
SP - 619
EP - 632
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 3
ER -
ID: 255066200