A novel fluorescence assay and catalytic properties of Crh1 and Crh2 yeast cell wall transglycosylases

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60162694%3AG44__%2F13%3A43875043" target="_blank" >RIV/60162694:G44__/13:43875043 - isvavai.cz</a>

Výsledek na webu

<a href="http://www.biochemj.org/bj/455/bj4550307.htm" target="_blank" >http://www.biochemj.org/bj/455/bj4550307.htm</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1042/BJ20130354" target="_blank" >10.1042/BJ20130354</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A novel fluorescence assay and catalytic properties of Crh1 and Crh2 yeast cell wall transglycosylases

Popis výsledku v původním jazyce

The mechanical properties of fungal cell walls are largely determined by composition and mutual cross-linking of their macromolecular components. Previous work showed that the Crh proteins are required for the formation of cross-links between chitin andglucan at the Saccharomyces cerevisiae cell wall. In the present study, the proteins encoded by CRH1 and CRH2 were heterologously expressed in Pichia pastoris and a sensitive fluorescence in vitro soluble assay was devised for determination of their transglycosylating activities. Both proteins act as chitin transglycosylases; they use soluble chitin derivatives, such as carboxymethyl chitin, glycol-chitin and/or N-acetyl chito-oligosaccharides of DP (degree of polymerization) >= 5 as the oligoglycosyl donors, and oligosaccharides derived from chitin, beta-(1,3)-glucan (laminarin) and beta-(1,6)-glucan (pustulan), fluorescently labelled with sulforhodamine or FITC as acceptors. The minimal number of intact hexopyranose units required byC

Název v anglickém jazyce

A novel fluorescence assay and catalytic properties of Crh1 and Crh2 yeast cell wall transglycosylases

Popis výsledku anglicky

The mechanical properties of fungal cell walls are largely determined by composition and mutual cross-linking of their macromolecular components. Previous work showed that the Crh proteins are required for the formation of cross-links between chitin andglucan at the Saccharomyces cerevisiae cell wall. In the present study, the proteins encoded by CRH1 and CRH2 were heterologously expressed in Pichia pastoris and a sensitive fluorescence in vitro soluble assay was devised for determination of their transglycosylating activities. Both proteins act as chitin transglycosylases; they use soluble chitin derivatives, such as carboxymethyl chitin, glycol-chitin and/or N-acetyl chito-oligosaccharides of DP (degree of polymerization) >= 5 as the oligoglycosyl donors, and oligosaccharides derived from chitin, beta-(1,3)-glucan (laminarin) and beta-(1,6)-glucan (pustulan), fluorescently labelled with sulforhodamine or FITC as acceptors. The minimal number of intact hexopyranose units required byC

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

—

Návaznosti

V - Vyzkumna aktivita podporovana z jinych verejnych zdroju

Rok uplatnění

2013

Kód důvěrnosti údajů

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

Název periodika

Biochemical Journal

ISSN

0264-6021

e-ISSN

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Svazek periodika

455

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

12

Strana od-do

307-318

Kód UT WoS článku

000326664100005

EID výsledku v databázi Scopus

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