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Start Over Descriptor "N-Terminal Acetyltransferase D" Publisher mdpi
2 results on '"N-Terminal Acetyltransferase D"'

1. Development of A Continuous Fluorescence-Based Assay for N-Terminal Acetyltransferase D

Author

Yi-Hsun Ho, Rong Huang, and Lan Chen

Subjects
High-throughput screening, Coenzyme A, N-terminal acetyltransferase D, Pilot Projects, high-throughput screening, Catalysis, Cofactor, Article, Fluorescence, Inorganic Chemistry, lcsh:Chemistry, Histone H4, Histones, N-Terminal Acetyltransferase D, chemistry.chemical_compound, Humans, Physical and Theoretical Chemistry, Molecular Biology, lcsh:QH301-705.5, Spectroscopy, Enzyme Assays, Fluorescent Dyes, biology, Chemistry, Organic Chemistry, fluorescence assay, Reproducibility of Results, Acetyltransferases, Acetylation, General Medicine, Computer Science Applications, High-Throughput Screening Assays, lcsh:Biology (General), lcsh:QD1-999, Biochemistry, Nat, Acetyltransferase, biology.protein, acetyltransferase inhibitors
Abstract

N-terminal acetylation catalyzed by N-terminal acetyltransferases (NATs) has various biological functions in protein regulation. N-terminal acetyltransferase D (NatD) is one of the most specific NAT with only histone H4 and H2A proteins as the known substrates. Dysregulation of NatD has been implicated in colorectal and lung cancer progression, implying its therapeutic potential in cancers. However, there is no reported inhibitor for NatD yet. To facilitate the discovery of small-molecule NatD inhibitors, we report the development of a fluorescence-based acetyltransferase assay in 384-well high-throughput screening (HTS) format through monitoring the formation of coenzyme A. The fluorescent signal is generated from the adduct in the reaction between coenzyme A and fluorescent probe ThioGlo4. The assay exhibited a Z&rsquo, factor of 0.77 and a coefficient of variation of 6%, indicating it is a robust assay for HTS. A pilot screen of 1280 pharmacologically active compounds and subsequent validation identified two hits, confirming the application of this fluorescence assay in HTS.

Published
2021

2. Development of A Continuous Fluorescence-Based Assay for N -Terminal Acetyltransferase D.

Author

Ho YH, Chen L, and Huang R

Subjects
Acetylation, Humans, Pilot Projects, Reproducibility of Results, Enzyme Assays methods, Fluorescence, Fluorescent Dyes chemistry, High-Throughput Screening Assays methods, Histones metabolism, N-Terminal Acetyltransferase D metabolism
Abstract

N -terminal acetylation catalyzed by N -terminal acetyltransferases (NATs) has various biological functions in protein regulation. N -terminal acetyltransferase D (NatD) is one of the most specific NAT with only histone H4 and H2A proteins as the known substrates. Dysregulation of NatD has been implicated in colorectal and lung cancer progression, implying its therapeutic potential in cancers. However, there is no reported inhibitor for NatD yet. To facilitate the discovery of small-molecule NatD inhibitors, we report the development of a fluorescence-based acetyltransferase assay in 384-well high-throughput screening (HTS) format through monitoring the formation of coenzyme A. The fluorescent signal is generated from the adduct in the reaction between coenzyme A and fluorescent probe ThioGlo4. The assay exhibited a Z'-factor of 0.77 and a coefficient of variation of 6%, indicating it is a robust assay for HTS. A pilot screen of 1280 pharmacologically active compounds and subsequent validation identified two hits, confirming the application of this fluorescence assay in HTS.

Published
2021
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