Diversity of serine hydrolase activities of unchallenged and botrytis-infected Arabidopsis thaliana.
Activity-based protein profiling is a powerful method to display enzyme activities in proteomes and provides crucial information on enzyme activity rather than protein or transcript abundance. We applied activity-based protein profiling using fluorophosphonate-based probes to display the activities of Ser hydrolases in the model plant Arabidopsis thaliana. Multidimensional protein identification technology and in-gel analysis of fluorophosphonate-labeled leaf extracts revealed over 50 Ser hydrolases, including dozens of proteases, esterases, and lipases, representing over 10 different enzyme families. Except for some well characterized Ser hydrolases like subtilases TPP2 and ARA12, prolyl oligopeptidase acylamino acid-releasing enzyme, serine carboxypeptidase-like SNG1 and BRS1, carboxylesterase-like CXE12, methylesterases MES2 and MES3, and S-formylglutathione hydrolase, the majority of these serine hydrolases have not been described before. We studied transiently expressed SNG1 and investigated plants infected with the fungal pathogen Botrytis cinerea. Besides the down-regulation of several Arabidopsis Ser hydrolase activities during Botrytis infection, we detected the activities of Botrytis-derived cutinases and lipases, which are thought to contribute to pathogenicity.
Kaschani
Farnusch
F
Plant Chemetics laboratory, Max Planck Institute for Plant Breeding Research, Carl-von-Linné Weg 10, 50829 Cologne, Germany.
Gu
Christian
C
Niessen
Sherry
S
Hoover
Heather
H
Cravatt
Benjamin F
BF
van der Hoorn
Renier A L
RA
D000217:Acyltransferases / Q000378:metabolism; D016960:Agrobacterium tumefaciens; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000382:microbiology*; D029681:Arabidopsis Proteins / Q000378:metabolism; D020171:Botrytis / Q000502:physiology*; D054884:Host-Pathogen Interactions; D008969:Molecular Sequence Data; D063065:Organophosphonates / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D018515:Plant Leaves / Q000201:enzymology / Q000382:microbiology; D012697:Serine Endopeptidases / Q000032:analysis / Q000378:metabolism*; D014020:Tissue Extracts; D014026:Nicotiana / Q000378:metabolism
D029681:Arabidopsis Proteins; D063065:Organophosphonates; D010455:Peptides; D014020:Tissue Extracts; D000217:Acyltransferases; C416511:sinapoylglucose malate sinapoyltransferase; D012697:Serine Endopeptidases