title: Solubilized elastin substrate for continuous fluorimetric assay of kinetics of elastases.
abstract: Elastolysis is central to progression of emphysema and aortic aneurysms. Characterization of steady-state enzyme kinetics of elastolysis is fettered by the insolubility of mature elastin and the polydispersity of solubilized elastin. We prepared a fluor-tagged, 100-kDa fraction (fEln-100) from commercial α-elastin. It is soluble, less heterogeneous in mass, cross-linked like mature elastin, and likely to retain the capacity of α-elastin to self-assemble. fEln-100 has introduced the ability to compare quantitatively the apparent k(cat) and K(m) of elastases. For example, metalloelastase (MMP-12) displays higher apparent affinity for fEln-100, while MMP-2 displays faster catalytic turnover.
authors.affiliation: Department of Biochemistry, University of Missouri, Columbia, 65211, USA.
authors.forename: Mark O
authors.initials: MO
authors.lastname: Palmier
authors.forename: Yan G
authors.initials: YG
authors.lastname: Fulcher
authors.forename: Steven R
authors.initials: SR
authors.lastname: Van Doren
mesh_terms: D004549:Elastin / Q000737:chemistry / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods; D005470:Fluorometry / Q000379:methods*; D007700:Kinetics; D010196:Pancreatic Elastase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004549:Elastin; D010196:Pancreatic Elastase