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@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 a 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, roč.~410, č.~28, s.~7441-7448. ISSN~1618-2642. Dostupné z: https://dx.doi.org/10.1007/s00216-018-1354-6.
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