Reassessing enzyme kinetics: Considering protease-as-substrate interactions in proteolytic networks.

Abstract:

Enzymes are catalysts in biochemical reactions that, by definition, increase rates of reactions without being altered or destroyed. However, when that enzyme is a protease, a subclass of enzymes that hydrolyze other proteins, and that protease is in a multiprotease system, protease-as-substrate dynamics must be included, challenging assumptions of enzyme inertness, shifting kinetic predictions of that system. Protease-on-protease inactivating hydrolysis can alter predicted protease concentrations used to determine pharmaceutical dosing strategies. Cysteine cathepsins are proteases capable of cathepsin cannibalism, where one cathepsin hydrolyzes another with substrate present, and misunderstanding of these dynamics may cause miscalculations of multiple proteases working in one proteolytic network of interactions occurring in a defined compartment. Once rates for individual protease-on-protease binding and catalysis are determined, proteolytic network dynamics can be explored using computational models of cooperative/competitive degradation by multiple proteases in one system, while simultaneously incorporating substrate cleavage. During parameter optimization, it was revealed that additional distraction reactions, where inactivated proteases become competitive inhibitors to remaining, active proteases, occurred, introducing another network reaction node. Taken together, improved predictions of substrate degradation in a multiple protease network were achieved after including reaction terms of autodigestion, inactivation, cannibalism, and distraction, altering kinetic considerations from other enzymatic systems, since enzyme can be lost to proteolytic degradation. We compiled and encoded these dynamics into an online platform (https://plattlab.shinyapps.io/catKLS/) for individual users to test hypotheses of specific perturbations to multiple cathepsins, substrates, and inhibitors, and predict shifts in proteolytic network reactions and system dynamics.

Profile Page: http://compmodelmatch.org/publications/10

PubMed ID: 31980525

Meetings: Finding Your Inner Modeler IV

Publication type: Journal

Journal: Proc Natl Acad Sci U S A

Citation: Proc Natl Acad Sci U S A. 2020 Feb 11;117(6):3307-3318. doi: 10.1073/pnas.1912207117. Epub 2020 Jan 24.

Date Published: 11th Feb 2020

Registered Mode: by PubMed ID

Authors: M. C. Ferrall-Fairbanks, C. A. Kieslich, M. O. Platt

help Creator
Creators
Not specified
Submitter
Activity

Views: 111

Created: 5th Aug 2021 at 17:44

Related items

Powered by
(v.1.10.3)
Copyright © 2008 - 2020 The University of Manchester and HITS gGmbH
2019-2021 University of Connecticut