Catalytic Mechanism of Processive GlfT2: Transition Path Sampling Investigation of Substrate Translocation

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F20%3A00117962" target="_blank" >RIV/00216224:14740/20:00117962 - isvavai.cz</a>

Result on the web

<a href="https://doi.org/10.1021/acsomega.0c01434" target="_blank" >https://doi.org/10.1021/acsomega.0c01434</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acsomega.0c01434" target="_blank" >10.1021/acsomega.0c01434</a>

Result language

angličtina

Original language name

Catalytic Mechanism of Processive GlfT2: Transition Path Sampling Investigation of Substrate Translocation

Original language description

We applied the transition path sampling (TPS) method to study the translocation step of the catalytic mechanism of galactofuranosyl transferase 2 (GlfT2). Using TPS in the field of enzymatic reactions is still relatively rare, and we show its effectiveness on this enzymatic system. We decipher an unknown mechanism of the translocation step and, thus, provide a complete understanding of the catalytic mechanism of GlfT2 at the atomistic level. The GlfT2 enzyme is involved in the formation of the mycobacterial cell wall and transfers galactofuranose (Galf) from UDP-Galf onto a growing acceptor Galf chain. The biosynthesis of the galactan chain is accomplished in a processive manner, with the growing acceptor substrate remaining bound to GlfT2. The glycosidic bond formed by GlfT2 between the two Gall residues alternates between beta-(1-6) and beta-(1-5) linkages. The translocation of the growing galactan between individual additions of Galf residues is crucial for the function of GlfT2. Analysis of unbiased trajectory ensembles revealed that the translocation proceeds differently depending on the glycosidic linkage between the last two Galf residues. We also showed that the protonation state of the catalytic residue Asp372 significantly influences the translocation. Approximate transition state structures and potential energy reaction barriers of the translocation process were determined. The calculated potential reaction barriers in the range of 6-14 kcal/mol show that the translocation process is not the rate-limiting step in galactan biosynthesis.

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

10400 - Chemical sciences

Project

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

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2020

Confidentiality

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

Name of the periodical

ACS Omega

ISSN

2470-1343

e-ISSN

—

Volume of the periodical

5

Issue of the periodical within the volume

34

Country of publishing house

US - UNITED STATES

Number of pages

11

Pages from-to

21374-21384

UT code for WoS article

000568645300006

EID of the result in the Scopus database

2-s2.0-85091020953