title: Inhibitor Discovery by Convolution ABPP.
abstract: Activity-based protein profiling (ABPP) has emerged as a powerful proteomic approach to study the active proteins in their native environment by using chemical probes that label active site residues in proteins. Traditionally, ABPP is classified as either comparative or competitive ABPP. In this protocol, we describe a simple method called convolution ABPP, which takes benefit from both the competitive and comparative ABPP. Convolution ABPP allows one to detect if a reduced signal observed during comparative ABPP could be due to the presence of inhibitors. In convolution ABPP, the proteomes are analyzed by comparing labeling intensities in two mixed proteomes that were labeled either before or after mixing. A reduction of labeling in the mix-and-label sample when compared to the label-and-mix sample indicates the presence of an inhibitor excess in one of the proteomes. This method is broadly applicable to detect inhibitors in proteomes against any proteome containing protein activities of interest. As a proof of concept, we applied convolution ABPP to analyze secreted proteomes from Pseudomonas syringae-infected Nicotiana benthamiana leaves to display the presence of a beta-galactosidase inhibitor.
authors: The Plant Chemetics Laboratory, Max Planck Institute for Plant Breeding Research, Carl-von-Linne Weg 10, 50829, Cologne, Germany.|The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK.
authors: Balakumaran
authors: B
authors: Chandrasekar
authors: The Plant Chemetics Laboratory, Max Planck Institute for Plant Breeding Research, Carl-von-Linne Weg 10, 50829, Cologne, Germany.|The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK.
authors: Tram Ngoc
authors: TN
authors: Hong
authors: The Plant Chemetics Laboratory, Max Planck Institute for Plant Breeding Research, Carl-von-Linne Weg 10, 50829, Cologne, Germany. renier.vanderhoorn@plants.ox.ac.uk.
authors: Renier A L
authors: RA
authors: van der Hoorn
mesh_terms: D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D015335:Molecular Probes / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D044224:Pseudomonas syringae / Q000302:isolation & purification; D014026:Nicotiana / Q000737:chemistry; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors
chemical_list: D004791:Enzyme Inhibitors; D015335:Molecular Probes; D011506:Proteins; D001616:beta-Galactosidase
keywords: Activity-based protein profiling (ABPP); Beta-galactosidase; Convolution ABPP; Cyclophellitol-aziridine; Galactostatin; Nicotiana benthamiana; Pseudomonas syringae; Secreted proteomes