title: Activity-based protein profiling of hydrolytic enzymes induced by gibberellic acid in isolated aleurone layers of malting barley.
abstract: During barley germination, the aleurone layer secretes most of the enzymes required to degrade the endosperm, many of which are yet to be characterized. We used activity-based protein profiling (ABPP) to detect a range of active enzymes extracted from aleurone layers isolated from grains of a commercial malting barley variety incubated with or without gibberellic acid (GA). Enzymes found to be induced by GA were putative aleurains, cathepsin-B-like proteases and serine hydrolases. By using an inhibitory sugar panel, a specific active retaining β-glycosidase in the barley aleurone was identified as a putative xylanase. Our results show that ABPP can be used rapidly to identify a variety of active enzyme isoforms in cereal aleurone without the need for enzyme purification.
authors.affiliation: Faculty of Agriculture and Environment, Plant Breeding Institute, University of Sydney, Eveleigh, Australia.
authors.forename: Sergio N
authors.initials: SN
authors.lastname: Daneri-Castro
authors.affiliation: The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, UK.
authors.forename: Balakumaran
authors.initials: B
authors.lastname: Chandrasekar
authors.affiliation: The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, UK.
authors.forename: Friederike M
authors.initials: FM
authors.lastname: Grosse-Holz
authors.affiliation: The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, UK.
authors.forename: Renier A L
authors.initials: RA
authors.lastname: van der Hoorn
authors.affiliation: Faculty of Agriculture and Environment, Plant Breeding Institute, University of Sydney, Eveleigh, Australia.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Roberts
mesh_terms: D000040:Abscisic Acid; D017209:Apoptosis / Q000187:drug effects; D056625:Endosperm / Q000201:enzymology* / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D018525:Germination / Q000235:genetics; D005875:Gibberellins / Q000378:metabolism* / Q000494:pharmacology; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D005875:Gibberellins; D010940:Plant Proteins; D017382:Reactive Oxygen Species; D000040:Abscisic Acid; C007842:gibberellic acid
keywords: activity-based protein profiling; aleurone layer; barley