New bioluminogenic substrates for monoamine oxidase assays.
Novel bioluminogenic substrates were designed for probing monoamine oxidase (MAO) activity based on a simple and effective beta-elimination strategy. By modifying the amino group and the central core of luciferin derivatives, we have developed a series of substrates useful for assays of MAO A or B, or both. One of these substrates, exhibiting low Km values and high signal-to-background ratios with both isozymes, was shown to accurately measure the Ki values of known MAO inhibitors. This substrate is a key component in the development of a highly sensitive homogeneous MAO assay for high-throughput screening (HTS) of compounds in drug discovery and for monitoring MAO activity in complex biological systems. This design strategy should be applicable to fluorogenic MAO substrates and could broaden the structural requirements of substrates for other enzyme assays.
Zhou
Wenhui
W
Promega Biosciences, Inc., 277 Granada Drive, San Luis Obispo, California 93401, USA. wenhui.zhou@promega.com
Valley
Michael P
MP
Shultz
John
J
Hawkins
Erika M
EM
Bernad
Laurent
L
Good
Troy
T
Good
Dave
D
Riss
Terry L
TL
Klaubert
Dieter H
DH
Wood
Keith V
KV
D049454:Firefly Luciferin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D007700:Kinetics; D049408:Luminescent Agents / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D008995:Monoamine Oxidase / Q000032:analysis* / Q000378:metabolism; D010084:Oxidation-Reduction; D013379:Substrate Specificity
D007527:Isoenzymes; D049408:Luminescent Agents; D049454:Firefly Luciferin; D008995:Monoamine Oxidase