Active Site Complementation and Hexameric Arrangement in the GH Family 29: A Structure-Function Study of α-L-Fucosidase Isoenzyme 1 from Paenibacillus thiaminolyticus.

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F19%3A43918196" target="_blank" >RIV/60461373:22330/19:43918196 - 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 α-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 α-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

—

Návaznosti

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

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

15

Strana od-do

59-73

Kód UT WoS článku

000462551800005

EID výsledku v databázi Scopus

2-s2.0-85058766539