使用荧光底物测量蛋白酶活性

Measurement of Protease Activities Using Fluorogenic Substrates.

作者信息

Santamaria Salvatore, Nagase Hideaki

机构信息

Centre for Haematology, Imperial College London, London, UK.

Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK.

出版信息

Methods Mol Biol. 2018;1731:107-122. doi: 10.1007/978-1-4939-7595-2_11.

DOI:10.1007/978-1-4939-7595-2_11

PMID:29318548

Abstract

Matrix metalloproteinases and the related metalloproteases are implicated in cancer progression. They are endopeptidases that require several defined amino acid residues in both N-terminal and C-terminal sides of the scissile bond. Fluorogenic Förster resonance energy transfer (FRET) substrates that harbor a fluorophore and a quencher on opposite sides of the scissile bond are conveniently used to measure their activities. In this chapter, we describe the principle of FRET substrates and how to use them to measure activities and kinetic parameters of endopeptidases.

摘要

基质金属蛋白酶及相关金属蛋白酶与癌症进展有关。它们是内肽酶，在可裂解键的N端和C端两侧都需要几个特定的氨基酸残基。在可裂解键两侧带有荧光团和猝灭剂的荧光共振能量转移（FRET）底物可方便地用于测量它们的活性。在本章中，我们描述了FRET底物的原理以及如何使用它们来测量内肽酶的活性和动力学参数。