Catalytic by-product formation and ligand binding by ribulose bisphosphate carboxylases from different phylogenies.
During catalysis, all Rubisco (D-ribulose-1,5-bisphosphate carboxylase/oxygenase) enzymes produce traces of several by-products. Some of these by-products are released slowly from the active site of Rubisco from higher plants, thus progressively inhibiting turnover. Prompted by observations that Form I Rubisco enzymes from cyanobacteria and red algae, and the Form II Rubisco enzyme from bacteria, do not show inhibition over time, the production and binding of catalytic by-products was measured to ascertain the underlying differences. In the present study we show that the Form IB Rubisco from the cyanobacterium Synechococcus PCC6301, the Form ID enzyme from the red alga Galdieria sulfuraria and the low-specificity Form II type from the bacterium Rhodospirillum rubrum all catalyse formation of by-products to varying degrees; however, the by-products are not inhibitory under substrate-saturated conditions. Study of the binding and release of phosphorylated analogues of the substrate or reaction intermediates revealed diverse strategies for avoiding inhibition. Rubisco from Synechococcus and R. rubrum have an increased rate of inhibitor release. G. sulfuraria Rubisco releases inhibitors very slowly, but has an increased binding constant and maintains the enzyme in an activated state. These strategies may provide information about enzyme dynamics, and the degree of enzyme flexibility. Our observations also illustrate the phylogenetic diversity of mechanisms for regulating Rubisco and raise questions about whether an activase-like mechanism should be expected outside the green-algal/higher-plant lineage.
Pearce
F Grant
FG
School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8020, New Zealand. grant.pearce@canterbury.ac.nz
D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D002384:Catalysis; D005988:Glyceric Acids / Q000378:metabolism; D008024:Ligands; D010755:Organophosphates / Q000378:metabolism; D010084:Oxidation-Reduction; D010428:Pentosephosphates / Q000378:metabolism / Q000494:pharmacology; D010802:Phylogeny; D010940:Plant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D011773:Pyruvates / Q000378:metabolism; D000461:Rhodophyta / Q000201:enzymology*; D012247:Rhodospirillum rubrum / Q000201:enzymology*; D012273:Ribulose-Bisphosphate Carboxylase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D012274:Ribulosephosphates / Q000378:metabolism*; D013045:Species Specificity; D018724:Spinacia oleracea / Q000201:enzymology; D013402:Sugar Alcohols / Q000378:metabolism / Q000494:pharmacology; D013403:Sugar Phosphates / Q000378:metabolism; D046940:Synechococcus / Q000201:enzymology*; D014026:Nicotiana / Q000201:enzymology*
C095598:2-carboxytetritol 1,4-bisphosphate; D001426:Bacterial Proteins; D005988:Glyceric Acids; D008024:Ligands; D010755:Organophosphates; D010428:Pentosephosphates; D010940:Plant Proteins; D011773:Pyruvates; D012274:Ribulosephosphates; D013402:Sugar Alcohols; D013403:Sugar Phosphates; C093484:glycero-2,3-pentodiulose 1,5-bisphosphate; C036130:xylulose 1,5-bisphosphate; C001933:ribulose-1,5 diphosphate; C032687:2-carboxyarabinitol 1,5-biphosphate; D012273:Ribulose-Bisphosphate Carboxylase