Reactive oxygen species affect ATP hydrolysis by targeting a highly conserved amino acid cluster in the thylakoid ATP synthase γ subunit.
The vast majority of organisms produce ATP by a membrane-bound rotating protein complex, termed F-ATP synthase. In chloroplasts, the corresponding enzyme generates ATP by using a transmembrane proton gradient generated during photosynthesis, a process releasing high amounts of molecular oxygen as a natural byproduct. Due to its chemical properties, oxygen can be reduced incompletely which generates several highly reactive oxygen species (ROS) that are able to oxidize a broad range of biomolecules. In extension to previous studies it could be shown that ROS dramatically decreased ATP synthesis in situ and affected the CF1 portion in vitro. A conserved cluster of three methionines and a cysteine on the chloroplast γ subunit could be identified by mass spectrometry to be oxidized by ROS. Analysis of amino acid substitutions in a hybrid F1 assembly system indicated that these residues were exclusive catalytic targets for hydrogen peroxide and singlet oxygen, although it could be deduced that additional unknown amino acid targets might be involved in the latter reaction. The cluster was tightly integrated in catalytic turnover since mutants varied in MgATPase rates, stimulation by sulfite and chloroplast-specific γ subunit redox-modulation. Some partial disruptions of the cluster by mutagenesis were dominant over others regarding their effects on catalysis and response to ROS.
Department of Plant Physiology, Justus-Liebig-University Giessen, Germany.
Felix
F
Buchert
Yvonne
Y
Schober
Andreas
A
Römpp
Mark L
ML
Richter
Christoph
C
Forreiter
D000255:Adenosine Triphosphate / Q000737:chemistry*; D000595:Amino Acid Sequence; D025222:Chloroplast Proton-Translocating ATPases / Q000737:chemistry / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D021122:Protein Subunits; D017382:Reactive Oxygen Species / Q000378:metabolism*; D026082:Singlet Oxygen / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013447:Sulfites / Q000494:pharmacology; D020524:Thylakoids / Q000201:enzymology*
D021122:Protein Subunits; D017382:Reactive Oxygen Species; D013447:Sulfites; D026082:Singlet Oxygen; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D025222:Chloroplast Proton-Translocating ATPases