Identification of Protease Specificity Using Biotin-Labeled Substrates
Hiroyuki Yamamoto1, *, Syota Saito1, Yoshikazu Sawaguchi2, Michio Kimura1
Identifiers and Pagination:Year: 2017
First Page: 27
Last Page: 35
Publisher ID: TOBIOCJ-11-27
Article History:Received Date: 19/08/2016
Revision Received Date: 14/02/2017
Acceptance Date: 17/03/2017
Electronic publication date: 21/04/2017
Collection year: 2017
open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Proteolysis constitutes a major post-translational modification. For example, proteases regulate the activation or inactivation of various proteins, such as enzymes, growth factors, and peptide hormones. Proteases have substrate specificity, and protease expression regulates the specific and regional activation or inactivation of several functional proteins.
We demonstrate a novel method for determining protease specificity through the use of MALDI-TOF mass spectrometry with biotin-labeled substrates.
This method was able to determine the specificity of TPCK-trypsin, V8 protease, elastase and cyanogen bromide cleavage, and the results were similar to previous reports. In addition, the method can be used to measure crude samples, such as tumor extracts.
We demonstrated that this method could identify protease specificity after simple processing, even for crude samples.