Collection Citation: Cristina Rodrigues Gabriel Sales, Anabela Silva, Elizabete Carmo-Silva 2020. Protocols from Sales et al. (2020) Rubisco activity: challenges and opportunities of NADH-linked microtiter plate-based and 14C-based assays. protocols.io https://dx.doi.org/10.17504/protocols.io.bf8djrs6
Manuscript citation:
Sales CRG, Silva AB, Carmo-Silva E. 2020. Measuring Rubisco activity: challenges and opportunities of NADH-linked microtiter plate-based and 14C-based assays. Journal of Experimental Botany, https://doi.org/10.1093/jxb/eraa289
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
This collection of protocols includes four assays for measuring Rubisco activity that are used and discussed in the manuscript "Rubisco activity: challenges and opportunities of NADH-linked microtiter plate-based and 14C-based assays" (Sales et al. 2020).
Three of the protocols are NADH-linked microtiter plate-based assays for Rubisco activity that use alternative coupling enzymes:
1. GAPDH-GlyPDH, glyceraldehyde-3-phosphate-dehydrogenase and glycerolphosphate-dehydrogenase;
2. PEPC-MDH, phosphoenolpyruvate-carboxylase and malate-dehydrogenase;
3. PK-LDH, pyruvate-kinase and lactate-dehydrogenase.
The radiometric assay for Rubisco activity measures the incorporation of 14CO2 into 3-phosphoglycerate (3-PGA).
The fifth protocol describes the purification of 2,3-bisphosphate-dependent phosphoglycerate mutase (dPGM), which is required for the NADH-linked microtiter plate-based assays 2 (PEPC-MDH) and 3 (PK-LDH).
Guidelines
Please, check each protocols for detailed guidelines.
