title: Detection of endogenous matrix metalloprotease-12 active form with a novel broad spectrum activity-based probe.
abstract: Matrix metalloproteases (MMPs) have attracted considerable attention as critical mediators of pathological tissue remodeling processes. However it remains an unresolved challenge to detect their active forms in biological samples. To prove the efficacy of a recently developed MMP activity-based probe, we examined the content in MMP active forms of bronchoalveolar lavage fluids (BALf) from male C57BL/6 mice exposed to ultrafine carbon black nanoparticles, a model of chronic obstructive pulmonary disease. This probe was shown to label proteins, mostly expressed in BALf of mice exposed to nanoparticles. Using competition assays with a selective MMP-12 inhibitor as well as MMP-12 knock-out mice, one of these proteins was identified as the active form of the catalytic domain of MMP-12. This new probe can detect the active form of MMP-12 down to a threshold of 1 fmol. Radioactive counting showed the concentration of the active form of MMP-12 to be around 1 fmol/μl in BALf from nanoparticle-treated mice. A less sensitive probe would therefore not have detected MMP-12. As the probe can detect other MMPs in the femtomolar range, it is a potentially powerful tool for monitoring the levels of MMP active forms in various diseases.
authors.affiliation: CEA (Commissariat à l'Energie Atomique), iBiTec-S, Service d'Ingénierie Moléculaire de Protéines (SIMOPRO), CE-Saclay, 91191 Gif /Yvette, Cedex, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Nury
authors.forename: Sarah
authors.initials: S
authors.lastname: Bregant
authors.forename: Bertrand
authors.initials: B
authors.lastname: Czarny
authors.forename: Fannely
authors.initials: F
authors.lastname: Berthon
authors.forename: Evelyne
authors.initials: E
authors.lastname: Cassar-Lajeunesse
authors.forename: Vincent
authors.initials: V
authors.lastname: Dive
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid; D020134:Catalytic Domain; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007249:Inflammation; D008297:Male; D053508:Matrix Metalloproteinase 12 / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008956:Models, Chemical; D053758:Nanoparticles; D029424:Pulmonary Disease, Chronic Obstructive / Q000378:metabolism; D000511:alpha-Macroglobulins / Q000378:metabolism
chemical_list: D015415:Biomarkers; D000511:alpha-Macroglobulins; D053508:Matrix Metalloproteinase 12