Design of an Activity-Based Probe for Human Neutrophil Elastase: Implementation of the Lossen Rearrangement To Induce Förster Resonance Energy Transfers.
Biochemistry
; 57(5): 742-752, 2018 02 06.
Article
em En
| MEDLINE
| ID: mdl-29286643
ABSTRACT
Human neutrophil elastase is an important regulator of the immune response and plays a role in host defense mechanisms and further physiological processes. The uncontrolled activity of this serine protease may cause severe tissue alterations and impair inflammatory states. The design of an activity-based probe for human neutrophil elastase reported herein relies on a sulfonyloxyphthalimide moiety as a new type of warhead that is linker-connected to a coumarin fluorophore. The inhibitory potency of the activity-based probe was assessed against several serine and cysteine proteases, and the selectivity for human neutrophil elastase (Ki = 6.85 nM) was determined. The adequate fluorescent tag of the probe allowed for the in-gel fluorescence detection of human neutrophil elastase in the low nanomolar range. The coumarin moiety and the anthranilic acid function of the probe, produced in the course of a Lossen rearrangement, were part of two different Förster resonance energy transfers.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Elastase de Leucócito
/
Transferência Ressonante de Energia de Fluorescência
/
Corantes Fluorescentes
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Alemanha