A label-free MALDI TOF MS-based method for studying the
kinetics and inhibitor screening of the Alzheimer’s disease
drug target beta-secretase

J 2018

A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase

MACHÁLKOVÁ, Markéta, Jan SCHEJBAL, Zdeněk GLATZ and Jan PREISLER

Basic information

Original name

A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase

Authors

MACHÁLKOVÁ, Markéta (203 Czech Republic, belonging to the institution), Jan SCHEJBAL (203 Czech Republic, belonging to the institution), Zdeněk GLATZ (203 Czech Republic, belonging to the institution) and Jan PREISLER (203 Czech Republic, guarantor, belonging to the institution)

Edition

Analytical and Bioanalytical Chemistry, Německo, Springer, 2018, 1618-2642

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.286

RIV identification code

RIV/00216224:14310/18:00101194

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1007/s00216-018-1354-6

UT WoS

000447989800017

Keywords in English

MALDI; mass spectrometry; BACE1; enzyme kinetics; enzyme inhibition; Alzheimer's disease

Abstract

V originále

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) is a well-established method with a unique set of qualities including sensitivity, minute sample consumption, and label-free detection, all of which are highly desired in enzyme assays. On the other hand, the application of MALDI TOF MS is usually limited by high concentrations of MS-incompatible compounds in the reaction mixture such as salts or organic solvents. Here, we introduce kinetic and inhibition studies of beta-secretase (BACE1), a key enzyme of the progression of Alzheimer’s disease. Compatibility of the enzyme assay with MALDI TOF MS was achieved, providing both a complex protocol including a desalting step designed for rigorous kinetic studies and a simple mix-and-measure protocol designed for high-throughput inhibitor screening. In comparison with fluorescent or colorimetric assays, MALDI TOF MS represents a sensitive, fast, and label-free technique with minimal sample preparation. In contrast to other MS-based methodological approaches typically used in drug discovery processes, such as a direct injection MS or MS-coupled liquid chromatography or capillary electrophoresis, MALDI TOF MS enables direct analysis and is a highly suitable approach for high-throughput screening. The method’s applicability is strongly supported by the high correlation of the acquired kinetic and inhibition parameters with data from the literature as well as from our previous research.

Links

GA16-06106S, research and development project

Name: Vysoce efektivní systém založený na kapilární elektroforéze pro screening inhibitorů beta-sekretázy jako terapeutického cíle pro Alzheimerovu chorobu (Acronym: Alzheimer)

Investor: Czech Science Foundation

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

MUNI/A/1278/2016, interní kód MU

Name: Podpora biochemického výzkumu v roce 2017

Investor: Masaryk University, Category A

MUNI/G/0974/2016, interní kód MU

Name: Optimalizace cílené terapie kolorektálního karcinomu ovlivňováním účinnosti penetrace a cytotoxicity regulátorů buněčných signalizací

Investor: Masaryk University, INTERDISCIPLINARY - Interdisciplinary research projects

Citovat

MACHÁLKOVÁ, Markéta, Jan SCHEJBAL, Zdeněk GLATZ and Jan PREISLER. A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase. Analytical and Bioanalytical Chemistry. Německo: Springer, 2018, vol. 410, No 28, p. 7441-7448. ISSN 1618-2642. Available from: https://dx.doi.org/10.1007/s00216-018-1354-6.

@article{1439537,
   author = {Machálková, Markéta and Schejbal, Jan and Glatz, Zdeněk and Preisler, Jan},
   article_location = {Německo},
   article_number = {28},
   doi = {http://dx.doi.org/10.1007/s00216-018-1354-6},
   keywords = {MALDI; mass spectrometry; BACE1; enzyme kinetics; enzyme inhibition; Alzheimer's disease},
   language = {eng},
   issn = {1618-2642},
   journal = {Analytical and Bioanalytical Chemistry},
   title = {A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase},
   url = {https://link.springer.com/article/10.1007%2Fs00216-018-1354-6},
   volume = {410},
   year = {2018}
}

TY  - JOUR
ID  - 1439537
AU  - Machálková, Markéta - Schejbal, Jan - Glatz, Zdeněk - Preisler, Jan
PY  - 2018
TI  - A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase
JF  - Analytical and Bioanalytical Chemistry
VL  - 410
IS  - 28
SP  - 7441-7448
EP  - 7441-7448
PB  - Springer
SN  - 16182642
KW  - MALDI
KW  - mass spectrometry
KW  - BACE1
KW  - enzyme kinetics
KW  - enzyme inhibition
KW  - Alzheimer's disease
UR  - https://link.springer.com/article/10.1007%2Fs00216-018-1354-6
N2  - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) is a well-established method with a unique set of qualities including sensitivity, minute sample consumption, and label-free detection, all of which are highly desired in enzyme assays. On the other hand, the application of MALDI TOF MS is usually limited by high concentrations of MS-incompatible compounds in the reaction mixture such as salts or organic solvents. Here, we introduce kinetic and inhibition studies of beta-secretase (BACE1), a key enzyme of the progression of Alzheimer’s disease. Compatibility of the enzyme assay with MALDI TOF MS was achieved, providing both a complex protocol including a desalting step designed for rigorous kinetic studies and a simple mix-and-measure protocol designed for high-throughput inhibitor screening. In comparison with fluorescent or colorimetric assays, MALDI TOF MS represents a sensitive, fast, and label-free technique with minimal sample preparation. In contrast to other MS-based methodological approaches typically used in drug discovery processes, such as a direct injection MS or MS-coupled liquid chromatography or capillary electrophoresis, MALDI TOF MS enables direct analysis and is a highly suitable approach for high-throughput screening. The method’s applicability is strongly supported by the high correlation of the acquired kinetic and inhibition parameters with data from the literature as well as from our previous research.
ER  -

MACHÁLKOVÁ, Markéta, Jan SCHEJBAL, Zdeněk GLATZ and Jan PREISLER. A label-free MALDI TOF MS-based method for studying the kinetics and inhibitor screening of the Alzheimer’s disease drug target beta-secretase. \textit{Analytical and Bioanalytical Chemistry}. Německo: Springer, 2018, vol.~410, No~28, p.~7441-7448. ISSN~1618-2642. Available from: https://dx.doi.org/10.1007/s00216-018-1354-6.

Detailed Information on Publication Record