scholarly journals In Vitro Activity Assays for MYST Histone Acetyltransferases and Adaptation for High-Throughput Inhibitor Screening

2016 ◽  
pp. 139-160 ◽  
Author(s):  
C.E. McCullough ◽  
R. Marmorstein
Keyword(s):  
High Throughput ◽  
Vitro Activity ◽  
Histone Acetyltransferases ◽  
In Vitro Activity ◽  
Inhibitor Screening

High-Throughput, Cell-Free, Liposome-Based Approach for Assessing In Vitro Activity of Lipid Kinases

2009 ◽  
Vol 14 (7) ◽  
pp. 838-844 ◽  
Author(s):  
Douglas J. Demian ◽  
Susan L. Clugston ◽  
Meta M. Foster ◽  
Lucia Rameh ◽  
Deborah Sarkes ◽  
...  
Keyword(s):  
High Throughput ◽  
Vitro Activity ◽  
Specific Substrate ◽  
In Vitro Activity ◽  
Lipid Kinase ◽  
Kinase Family ◽  
Large Numbers ◽  
Phosphatidylinositol Phosphate ◽  
Lipid Kinases

Lipid kinases are central players in lipid signaling pathways involved in inflammation, tumorigenesis, and metabolic syndrome. A number of these kinase targets have proven difficult to investigate in higher throughput cell-free assay systems. This challenge is partially due to specific substrate interaction requirements for several of the lipid kinase family members and the resulting incompatibility of these substrates with most established, homogeneous assay formats. Traditional, cell-free in vitro investigational methods for members of the lipid kinase family typically involve substrate incorporation of [γ-32P] and resolution of signal by thin-layer chromatography (TLC) and autoradiograph densitometry. This approach, although highly sensitive, does not lend itself to high-throughput testing of large numbers of small molecules (100 s to 1 MM+). The authors present the development and implementation of a fully synthetic, liposome-based assay for assessing in vitro activity of phosphatidylinositol-5-phosphate-4-kinase isoforms (PIP4KIIβ and α) in 2 commonly used homogeneous technologies. They have validated these assays through compound testing in both traditional TLC and radioactive filterplate approaches as well as binding validation using isothermic calorimetry. A directed library representing known kinase pharmacophores was screened against type IIβ phosphatidylinositol-phosphate kinase (PIPK) to identify small-molecule inhibitors. This assay system can be applied to other types and isoforms of PIPKs as well as a variety of other lipid kinase targets. ( Journal of Biomolecular Screening 2009:838-844)

In Vitro Activity of Phytocannabinoids from High Potency Cannabis sativa

Planta Medica ◽  
2012 ◽  
Vol 78 (05) ◽  
Author(s):  
A Husni ◽  
S Ross ◽  
O Dale ◽  
C Gemelli ◽  
G Ma ◽  
...  
Keyword(s):  
Cannabis Sativa ◽  
Vitro Activity ◽  
In Vitro Activity ◽  
High Potency
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