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Characterization of a photosynthesizing reconstituted spinach chloroplast preparation: regulation by primer, adenylates, ferredoxin, and pyridine nucleotides

Journal Article · · Plant Physiol.; (United States)
DOI:https://doi.org/10.1104/pp.69.1.179· OSTI ID:5799740

A particulate preparation (MgP) capable of photosynthetic CO/sub 2/ assimilation without the addition of stromal protein was obtained by rupturing whole spinach (Spinacia oleracea var. America) chloroplasts in 15 millimolar MgCl/sub 2/ buffered with Tricine at pH 8.5. This CO/sub 2/ assimilation was dependent upon light, inorganic phosphate, ferredoxin, ADP, NAD or NADP, and primer. Excepting glycolate, the products of CO/sub 2/ fixation by MgP were similar to those found with whole chloroplasts. Glycerate-3-phosphate (PGA), fructose-1,6-bisphosphate (FBP), and ribose-5-phosphate (R5P) but not fructose-6-P (F6P) functioned as primers. Concentrations of PGA and FBP but not of R5P higher than 2 millimolar where inhibitory to CO/sub 2/ fixation. A lag of CO/sub 2/ fixation was observed with PGA and FBP but not with R5P. This lag as well as inhibition by NADP, ADP, and ATP in the FBP-primed preparation was eliminated by an equimolar mixture of FBP plus F6P indicating FBPase as the sensitive site. NADP, ADP, and ATP also blocked CO/sub 2/ fixation by the PGA-fortified preparation but inhibition was even more sensitive than that observed when FBP was added. Inhibition by AMP in the PGA and FBP-primed preparations was not affected by the addition of F6P. When R5P was the starting primer, inhibition of CO/sub 2/ fixation was relatively insensitive to the adenylates and NADP. In contrast to the parent whole chloroplast, CO/sub 2/ fixation by MgP was insensitive to high (5 millimolar) inorganic phosphate. Depending upon the ferredoxin concentration, NAD was as effective as NADP in supporting CO/sub 2/ fixation.

Research Organization:
Brandeis Univ., Waltham, MA
OSTI ID:
5799740
Journal Information:
Plant Physiol.; (United States), Journal Name: Plant Physiol.; (United States) Vol. 69:1; ISSN PLPHA
Country of Publication:
United States
Language:
English