Abstract
Photorespiration is an essential metabolic process in plants occurring via the oxygenase reaction of RuBisCO. In order to understand this process, it is essential to determine the amounts of intermediates involved. For this purpose we combined mass spectrometry-based approaches and the use of authentic standards for the quantification of photorespiratory intermediates. Here we describe protocols for the extraction and quantification of 2-phosphoglycolate (2PG) by LC-MS/MS and serine, glycine, glycolate, hydroxypyruvate, glyoxylate, and glycerate by GC-MS.
Keywords:
Authentic standards; GC-MS; LC-MS/MS; Metabolite quantification; Photorespiratory intermediates.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Arabidopsis / chemistry
-
Arabidopsis / metabolism*
-
Chromatography, Liquid / instrumentation
-
Chromatography, Liquid / methods
-
Gas Chromatography-Mass Spectrometry / instrumentation
-
Gas Chromatography-Mass Spectrometry / methods
-
Glyceric Acids / isolation & purification
-
Glyceric Acids / metabolism
-
Glycine / isolation & purification
-
Glycine / metabolism
-
Glycolates / isolation & purification
-
Glycolates / metabolism
-
Glyoxylates / isolation & purification
-
Glyoxylates / metabolism
-
Mass Spectrometry / instrumentation
-
Mass Spectrometry / methods*
-
Oxidation-Reduction
-
Oxygen Consumption / physiology*
-
Photosynthesis / physiology*
-
Plant Leaves / chemistry
-
Plant Leaves / metabolism*
-
Pyruvates / isolation & purification
-
Pyruvates / metabolism
-
Reference Standards
-
Ribulose-Bisphosphate Carboxylase / metabolism*
-
Serine / isolation & purification
-
Serine / metabolism
Substances
-
Glyceric Acids
-
Glycolates
-
Glyoxylates
-
Pyruvates
-
glycolic acid
-
Serine
-
glyceric acid
-
hydroxypyruvic acid
-
Ribulose-Bisphosphate Carboxylase
-
phosphoglycolate
-
glyoxylic acid
-
Glycine