Active site complementation and hexameric arrangement in the GH family 29, a structure-function study of alpha-l-fucosidase isoenzyme 1 from Paenibacillus thiaminolyticus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F19%3A00510112" target="_blank" >RIV/86652036:_____/19:00510112 - isvavai.cz</a>

Výsledek na webu

<a href="https://academic.oup.com/glycob/article/29/1/59/5078562" target="_blank" >https://academic.oup.com/glycob/article/29/1/59/5078562</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1093/glycob/cwy078" target="_blank" >10.1093/glycob/cwy078</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Active site complementation and hexameric arrangement in the GH family 29, a structure-function study of alpha-l-fucosidase isoenzyme 1 from Paenibacillus thiaminolyticus

Popis výsledku v původním jazyce

alpha-l-Fucosidase isoenzyme 1 from bacterium Paenibacillus thiaminolyticus is a member of the glycoside hydrolase family GH29 capable of cleaving l-fucose from nonreducing termini of oligosaccharides and glycoconjugates. Here we present the first crystal structure of this protein revealing a novel quaternary state within this family. The protein is in a unique hexameric assembly revealing the first observed case of active site complementation by a residue from an adjacent monomer in this family. Mutation of the complementing tryptophan residue caused changes in the catalytic properties including a shift of the pH optimum, a change of affinity to an artificial chromogenic substrate and a decreased reaction rate for a natural substrate. The wild-type enzyme was active on most of the tested naturally occurring oligosaccharides and capable of transglycosylation on a variety of acceptor molecules, including saccharides, alcohols or chromogenic substrates. Mutation of the complementing residue changed neither substrate specificity nor the preference for the type of transglycosylation acceptor molecule, however, the yields of the reactions were lower in both cases. Maltose molecules bound to the enzyme in the crystal structure identified surface carbohydrate-binding sites, possibly participating in binding of larger oligosaccharides.

Název v anglickém jazyce

Active site complementation and hexameric arrangement in the GH family 29, a structure-function study of alpha-l-fucosidase isoenzyme 1 from Paenibacillus thiaminolyticus

Popis výsledku anglicky

alpha-l-Fucosidase isoenzyme 1 from bacterium Paenibacillus thiaminolyticus is a member of the glycoside hydrolase family GH29 capable of cleaving l-fucose from nonreducing termini of oligosaccharides and glycoconjugates. Here we present the first crystal structure of this protein revealing a novel quaternary state within this family. The protein is in a unique hexameric assembly revealing the first observed case of active site complementation by a residue from an adjacent monomer in this family. Mutation of the complementing tryptophan residue caused changes in the catalytic properties including a shift of the pH optimum, a change of affinity to an artificial chromogenic substrate and a decreased reaction rate for a natural substrate. The wild-type enzyme was active on most of the tested naturally occurring oligosaccharides and capable of transglycosylation on a variety of acceptor molecules, including saccharides, alcohols or chromogenic substrates. Mutation of the complementing residue changed neither substrate specificity nor the preference for the type of transglycosylation acceptor molecule, however, the yields of the reactions were lower in both cases. Maltose molecules bound to the enzyme in the crystal structure identified surface carbohydrate-binding sites, possibly participating in binding of larger oligosaccharides.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2019

Kód důvěrnosti údajů

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

Název periodika

Glycobiology

ISSN

0959-6658

e-ISSN

—

Svazek periodika

29

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

24

Strana od-do

59-73

Kód UT WoS článku

000462551800005

EID výsledku v databázi Scopus

—