Proteases Immobilization for In Situ Time-Limited Proteolysis on MALDI Chips

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F19%3A00518684" target="_blank" >RIV/61388971:_____/19:00518684 - isvavai.cz</a>

Alternative codes found

RIV/00216208:11310/19:10402940

Result on the web

<a href="https://www.mdpi.com/2073-4344/9/10/833" target="_blank" >https://www.mdpi.com/2073-4344/9/10/833</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/catal9100833" target="_blank" >10.3390/catal9100833</a>

Result language

angličtina

Original language name

Proteases Immobilization for In Situ Time-Limited Proteolysis on MALDI Chips

Original language description

A large number of different enzyme immobilization techniques are used in the field of life sciences, clinical diagnostics, or biotechnology. Most of them are based on a chemically mediated formation of covalent bond between an enzyme and support material. The covalent bond formation is usually associated with changes of the enzymes' three-dimensional structure that can lead to reduction of enzyme activity. The present work demonstrates a potential of an ambient ion-landing technique to effectively immobilize enzymes on conductive supports for direct matrix-assisted laser desorption/ionization (MALDI) mass spectrometry analyses of reaction products. Ambient ion landing is an electrospray-based technique allowing strong and stable noncovalent and nondestructive enzyme deposition onto conductive supports. Three serine proteolytic enzymes including trypsin, alpha-chymotrypsin, and subtilisin A were immobilized onto conductive indium tin oxide glass slides compatible with MALDI mass spectrometry. The functionalized MALDI chips were used for in situ time-limited proteolysis of proteins and protein-ligand complexes to monitor their structural changes under different conditions. The data from limited proteolysis using MALDI chips fits to known or predicted protein structures. The results show that functionalized MALDI chips are sensitive, robust, and fast and might be automated for general use in the field of structural biology.

Czech name

—

Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

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

2019

Confidentiality

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

Name of the periodical

Catalysts

ISSN

2073-4344

e-ISSN

—

Volume of the periodical

9

Issue of the periodical within the volume

10

Country of publishing house

CH - SWITZERLAND

Number of pages

11

Pages from-to

833

UT code for WoS article

000498266100049

EID of the result in the Scopus database

2-s2.0-85073535361