Aspartic Protease Nepenthesin-1 as a Tool for Digestion in Hydrogen/Deuterium Exchange Mass Spectrometry

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F14%3A10282688" target="_blank" >RIV/00216208:11310/14:10282688 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/14:00429336

Výsledek na webu

<a href="http://dx.doi.org/10.1021/ac404076j" target="_blank" >http://dx.doi.org/10.1021/ac404076j</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/ac404076j" target="_blank" >10.1021/ac404076j</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Aspartic Protease Nepenthesin-1 as a Tool for Digestion in Hydrogen/Deuterium Exchange Mass Spectrometry

Popis výsledku v původním jazyce

Hydrogen/deuterium exchange coupled to mass spectrometry (HXMS) utilizes enzymatic digestion of proteins to localize the information about altered exchange patterns in protein structure. The ability of the protease to produce small peptides and overlapping fragments and provide sufficient coverage of the protein sequence is essential for localizing regions of interest. Recently, it was shown that there is an interesting group of proteolytic enzymes from carnivorous pitcher plants of the genus Nepenthes.In this report, we describe successful immobilization and the use of one of these enzymes, nepenthesin-1, in HXMS workflow. In contrast to pepsin, it has different cleavage specificities, and despite its high inherent susceptibility to reducing and denaturing agents, it is very stable upon immobilization and withstands even high concentration of guanidine hydrochloride and reducing agents. We show that denaturing agents can alter digestion by reducing protease activity and/or substrate

Název v anglickém jazyce

Aspartic Protease Nepenthesin-1 as a Tool for Digestion in Hydrogen/Deuterium Exchange Mass Spectrometry

Popis výsledku anglicky

Hydrogen/deuterium exchange coupled to mass spectrometry (HXMS) utilizes enzymatic digestion of proteins to localize the information about altered exchange patterns in protein structure. The ability of the protease to produce small peptides and overlapping fragments and provide sufficient coverage of the protein sequence is essential for localizing regions of interest. Recently, it was shown that there is an interesting group of proteolytic enzymes from carnivorous pitcher plants of the genus Nepenthes.In this report, we describe successful immobilization and the use of one of these enzymes, nepenthesin-1, in HXMS workflow. In contrast to pepsin, it has different cleavage specificities, and despite its high inherent susceptibility to reducing and denaturing agents, it is very stable upon immobilization and withstands even high concentration of guanidine hydrochloride and reducing agents. We show that denaturing agents can alter digestion by reducing protease activity and/or substrate

Druh

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

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

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

Návaznosti

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

Rok uplatnění

2014

Kód důvěrnosti údajů

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

Název periodika

Analytical Chemistry

ISSN

0003-2700

e-ISSN

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Svazek periodika

86

Číslo periodika v rámci svazku

9

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

8

Strana od-do

4287-4294

Kód UT WoS článku

000335719900033

EID výsledku v databázi Scopus

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