Solubilized elastin substrate for continuous fluorimetric assay of kinetics of elastases.

Solubilized elastin substrate for continuous fluorimetric assay of kinetics of elastases.
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DOI:
10.1016/j.ab.2010.09.001
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发表时间:
2011-01-01
影响因子:
2.9
通讯作者:
Van Doren SR
Van Doren SR
中科院分区:
生物学4区
文献类型:
--
作者:
Palmier MO;Fulcher YG;Van Doren SR

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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 polydispersity of solubilized elastin. We prepared a fluor-tagged, 100 kDa fraction (fEln-100) from commercial α-elastin. It is soluble, less heterogenous 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 kcat and Km of elastases. For example, metalloelastase (MMP-12) displays higher apparent affinity for fEln-100, while MMP-2 displays faster catalytic turnover.
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