Structure of the Calvin-Benson-Bassham sedoheptulose-1,7-bisphosphatase from the model microalga <i>Chlamydomonas reinhardtii</i>.

Abstract

The Calvin-Benson-Bassham cycle (CBBC) performs carbon fixation in photosynthetic organisms. Among the eleven enzymes that participate in the pathway, sedoheptulose-1,7-bisphosphatase (SBPase) is expressed in photo-autotrophs and catalyzes the hydrolysis of sedoheptulose-1,7-bisphosphate (SBP) to sedoheptulose-7-phosphate (S7P). SBPase, along with nine other enzymes in the CBBC, contributes to the regeneration of ribulose-1,5-bisphosphate, the carbon-fixing co-substrate used by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The metabolic role of SBPase is restricted to the CBBC, and a recent study revealed that the three-dimensional structure of SBPase from the moss <i>Physcomitrium patens</i> was found to be similar to that of fructose-1,6-bisphosphatase (FBPase), an enzyme involved in both CBBC and neoglucogenesis. In this study we report the first structure of an SBPase from a chlorophyte, the model unicellular green microalga <i>Chlamydomonas reinhardtii</i>. By combining experimental and computational structural analyses, we describe the topology, conformations, and quaternary structure of <i>Chlamydomonas reinhardtii</i> SBPase (<i>Cr</i>SBPase). We identify active site residues and locate sites of redox- and phospho-post-translational modifications that contribute to enzymatic functions. Finally, we observe that <i>Cr</i>SBPase adopts distinct oligomeric states that may dynamically contribute to the control of its activity.