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New protocols to study photorespiration

Metaboactions contribute new protocols to the book "Photorespiration" in the Methods in Molecular Biology series

Members of the MetaboActions team at IPS2 have contributed to the book "Photorespiration" in the Methods in Molecular Biology series published by Springer. This book series was the first to introduce readily reproducible highly detailed step-by-step protocols for specific topics. Each protocol has an introductory overview, a list of materials and reagents, and a detailed procedure supported by numerous help notes offering tips, tricks and troubleshooting advice.


Photorespiration is an essential high flux metabolic pathway arising from the oxygenase activity of the most abundant leaf protein, Rubisco. It recycles toxic 2-phosphoglycolate to make useful 3-phosphoglycerate to fuel the Calvin cycle and the production of carbohydrates. This is an over-simplified view as the photorespiratory cycle interacts with several primary metabolic pathways including N-assimilation, amino acid biosynthesis, and energy metabolism. Since photorespiration is a high energy demanding process that competes with photosynthetic CO2 assimilation, it has become a target to improve crop yield. Indeed, photorespiration can reduce yields by 20-35%, and this is expected to increase due to climate change.


In this book, MetaboActions team members share their expertise in biochemistry, enzymology and NMR in three chapters that describe protocols they developed at IPS2. Two are dedicated to measuring 2-phosphoglycolate phosphatase activity using either NMR detection of glycolate or by spectrophotometry with a coupled reaction using recombinant glycolate oxidase. A third chapter describes the purification of recombinant N-terminal histidine-tagged Arabidopsis thaliana 2-phosphoglycolate phosphatase, glycolate oxidase and hydroxypyruvate reductase by affinity chromatography. These protocols are currently being used to study post-translational regulations of photorespiratory enzymes.

Book Cover
Book Cover