Deciphering Enzyme Mechanisms with Engineered Ancestors and Substrate Analogues

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F23%3A00079702" target="_blank" >RIV/00159816:_____/23:00079702 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/23:00132027

Result on the web

<a href="https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/cctc.202300745" target="_blank" >https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/cctc.202300745</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/cctc.202300745" target="_blank" >10.1002/cctc.202300745</a>

Result language

angličtina

Original language name

Deciphering Enzyme Mechanisms with Engineered Ancestors and Substrate Analogues

Original language description

Environmentally friendly industrial and biotech processes greatly benefit from enzyme-based technologies. Their use is often possible only when the enzyme-catalytic mechanism is thoroughly known. Thus, atomic-level knowledge of a Michaelis enzyme-substrate complex, revealing molecular details of substrate recognition and catalytic chemistry, is crucial for understanding and then rationally extending or improving enzyme-catalyzed reactions. However, many known enzymes sample huge protein conformational space, often preventing complete structural characterization by X-ray crystallography. Moreover, using a cognate substrate is problematic since its conversion into a reaction product in the presence of the enzyme will prevent the capture of the enzyme-substrate conformation in an activated state. Here, we outlined how to deal with such obstacles, focusing on the recent discovery of a Renilla-type bioluminescence reaction mechanism facilitated by a combination of engineered ancestral enzyme and the availability of a non-oxidizable luciferin analogue. The automated ancestral sequence reconstructions using FireProtASR provided a thermostable enzyme suited for structural studies, and a stable luciferin analogue azacoelenterazine provided a structurally cognate chemical incapable of catalyzed oxidation. We suggest that an analogous strategy can be applied to various enzymes with unknown catalytic mechanisms and poor crystallizability. Many known enzymes sample huge protein conformational space, hampering structural characterization by X-ray crystallography, and preventing thus the understanding of their catalytic mechanisms. In this review, we outline that the combination of reconstructed ancestral enzymes with unconvertible substrate analogues is becoming a powerful strategy to decipher the challenging mechanisms of enzyme catalysis.image

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10403 - Physical chemistry

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

—

Publication year

2023

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

ChemCatChem

ISSN

1867-3880

e-ISSN

1867-3899

Volume of the periodical

15

Issue of the periodical within the volume

19

Country of publishing house

DE - GERMANY

Number of pages

15

Pages from-to

—

UT code for WoS article

001060401600001

EID of the result in the Scopus database

—