Proton flux through the chloroplast ATP synthase is altered by cleavage of its gamma subunit.
Electron transport, the proton gradient and ATP synthesis were determined in thylakoids that had been briefly exposed to a low concentration of trypsin during illumination. This treatment cleaves the gamma subunit of the ATP synthase into two large fragments that remain associated with the enzyme. Higher rates of electron transport are required to generate a given value of the proton gradient in the trypsin-treated membranes than in control membranes, indicating that the treated membranes are proton leaky. Since venturicidin restores electron transport and the proton gradient to control levels, the proton leak is through the ATP synthase. Remarkably, the synthesis of ATP by the trypsin-treated membranes at saturating light intensities is only slightly inhibited even though the proton gradient is significantly lower in the treated thylakoids. ATP synthesis and the proton gradient were determined as a function of light intensity in control and trypsin-treated thylakoids. The trypsin-treated membranes synthesized ATP at lower values of the proton gradient than the control membranes. Cleavage of the gamma subunit abrogates inhibition of the activity of the chloroplast ATP synthase by the epsilon subunit. Our results suggest that overcoming inhibition by the epsilon subunit costs energy.
McCallum
Jeremy R
JR
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218, USA.
McCarty
Richard E
RE
D000255:Adenosine Triphosphate / Q000096:biosynthesis*; D002384:Catalysis; D025222:Chloroplast Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D004579:Electron Transport; D008027:Light; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011522:Protons; D018724:Spinacia oleracea / Q000201:enzymology; D020524:Thylakoids / Q000737:chemistry / Q000201:enzymology* / Q000528:radiation effects; D014357:Trypsin / Q000737:chemistry
D021122:Protein Subunits; D011522:Protons; D000255:Adenosine Triphosphate; D014357:Trypsin; D025222:Chloroplast Proton-Translocating ATPases