Premium
Small‐Molecule‐Induced and Cooperative Enzyme Assembly on a 14‐3‐3 Scaffold
Author(s) -
den Hamer Anniek,
Lemmens Lenne J. M.,
Nijenhuis Minke A. D.,
Ottmann Christian,
Merkx Maarten,
de Greef Tom F. A.,
Brunsveld Luc
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600631
Subject(s) - scaffold , scaffold protein , cooperativity , small molecule , chemistry , biophysics , protein–protein interaction , in vitro , signalling , nanotechnology , computational biology , microbiology and biotechnology , combinatorial chemistry , biochemistry , biology , computer science , signal transduction , materials science , database
Scaffold proteins regulate cell signalling by promoting the proximity of putative interaction partners. Although they are frequently applied in cellular settings, fundamental understanding of them in terms of, amongst other factors, quantitative parameters has been lagging behind. Here we present a scaffold protein platform that is based on the native 14‐3‐3 dimeric protein and is controllable through the action of a small‐molecule compound, thus permitting study in an in vitro setting and mathematical description. Robust small‐molecule regulation of caspase‐9 activity through induced dimerisation on the 14‐3‐3 scaffold was demonstrated. The individual parameters of this system were precisely determined and used to develop a mathematical model of the scaffolding concept. This model was used to elucidate the strong cooperativity of the enzyme activation mediated by the 14‐3‐3 scaffold. This work provides an entry point for the long‐needed quantitative insights into scaffold protein functioning and paves the way for the optimal use of reengineered 14‐3‐3 proteins as chemically inducible scaffolds in synthetic systems.