title: Labeling and enrichment of Arabidopsis thaliana matrix metalloproteases using an active-site directed, marimastat-based photoreactive probe.
abstract: Matrix metalloproteases (MMPs) are secreted or membrane-bound zinc-containing proteases that play diverse roles in development and immunity in plants and in tissue remodeling in animals. We developed a photoreactive probe based on the MMP inhibitor marimastat, conjugated to a 4-azidotetrafluorobenzoyl moiety as photoreactive group and biotin as detection or sorting function. The probe labels At2-MMP, At4-MMP, At5-MMP, and likely other plant MMPs in leaf extracts, as shown by transient At-MMP expression in Nicotiana benthamiana, protein blot, and LC-MS/MS analysis. This MMP probe is a valuable tool to study the post-translational status of MMPs during plant immunity and other MMP-regulated processes.
authors.affiliation: Bioorganic and Organic Chemistry, Department of Chemistry, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld, Germany.
authors.forename: Janina
authors.initials: J
authors.lastname: Lenger
authors.forename: Farnusch
authors.initials: F
authors.lastname: Kaschani
authors.forename: Thomas
authors.initials: T
authors.lastname: Lenz
authors.forename: Christian
authors.initials: C
authors.lastname: Dalhoff
authors.forename: Joji Grace
authors.initials: JG
authors.lastname: Villamor
authors.forename: Hubert
authors.initials: H
authors.lastname: Köster
authors.forename: Norbert
authors.initials: N
authors.lastname: Sewald
authors.forename: Renier A L
authors.initials: RA
authors.lastname: van der Hoorn
mesh_terms: D017360:Arabidopsis / Q000201:enzymology*; D020134:Catalytic Domain; D002851:Chromatography, High Pressure Liquid; D006877:Hydroxamic Acids / Q000737:chemistry*; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011480:Protease Inhibitors / Q000737:chemistry*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014466:Ultraviolet Rays
chemical_list: D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; D010940:Plant Proteins; D011480:Protease Inhibitors; D011994:Recombinant Proteins; C100342:marimastat; D020782:Matrix Metalloproteinases