Sulfide detoxification in plant mitochondria.
In contrast to animals, which release the signal molecule sulfide in small amounts from cysteine and its derivates, phototrophic eukaryotes generate sulfide as an essential intermediate of the sulfur assimilation pathway. Additionally, iron-sulfur cluster turnover and cyanide detoxification might contribute to the release of sulfide in mitochondria. However, sulfide is a potent inhibitor of cytochrome c oxidase in mitochondria. Thus, efficient sulfide detoxification mechanisms are required in mitochondria to ensure adequate energy production and consequently survival of the plant cell. Two enzymes have been recently described to catalyze sulfide detoxification in mitochondria of Arabidopsis thaliana, O-acetylserine(thiol)lyase C (OAS-TL C), and the sulfur dioxygenase (SDO) ethylmalonic encephalopathy protein 1 (ETHE1). Biochemical characterization of sulfide producing and consuming enzymes in mitochondria of plants is fundamental to understand the regulatory network that enables mitochondrial sulfide homeostasis under nonstressed and stressed conditions. In this chapter, we provide established protocols to determine the activity of the sulfide releasing enzyme β-cyanoalanine synthase as well as sulfide-consuming enzymes OAS-TL and SDO. Additionally, we describe a reliable and efficient method to purify OAS-TL proteins from plant material.
Centre for Organismal Studies Heidelberg, University of Heidelberg, Heidelberg, Germany.
Hannah
H
Birke
Institute for Plant Genetics, Leibniz University Hannover, Hannover, Germany.
Tatjana M
TM
Hildebrandt
Centre for Organismal Studies Heidelberg, University of Heidelberg, Heidelberg, Germany.
Markus
M
Wirtz
Centre for Organismal Studies Heidelberg, University of Heidelberg, Heidelberg, Germany. Electronic address: ruediger.hell@cos.uni-heidelberg.de.
Rüdiger
R
Hell
D017360:Arabidopsis / Q000737:chemistry / Q000201:enzymology; D029681:Arabidopsis Proteins / Q000302:isolation & purification / Q000378:metabolism*; D019757:Carbon-Oxygen Lyases / Q000302:isolation & purification / Q000378:metabolism*; D049308:Dioxygenases / Q000302:isolation & purification / Q000378:metabolism*; D057075:Enzyme Assays; D006862:Hydrogen Sulfide / Q000378:metabolism*; D007700:Kinetics; D008190:Lyases / Q000302:isolation & purification / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D051050:Serine O-Acetyltransferase / Q000737:chemistry
D029681:Arabidopsis Proteins; D049308:Dioxygenases; C000593849:ETHE1 protein, Arabidopsis; C477654:sulfur dioxygenase; D051050:Serine O-Acetyltransferase; C018725:O-acetylhomoserine (thiol)-lyase; D008190:Lyases; D019757:Carbon-Oxygen Lyases; C019955:beta-cyanoalanine synthase; D006862:Hydrogen Sulfide
Arabidopsis thaliana; Cytochrome c oxidase; ETHE1; Mitochondria; O-Acetylserine(thiol)lyase; Plant; SAT affinity purification; Sulfide toxicity; Sulfur dioxygenase; β-Cyanoalanine synthase