High Throughput Screen Identifies Small Molecule Effectors That Modulate Thin Filament Activation in Cardiac Muscle

2020 ◽  
Author(s):  
Priyanka Parijat ◽  
Laszlo Kondacs ◽  
Alexander Alexandrovich ◽  
Mathias Gautel ◽  
Alexander J. A. Cobb ◽  
...  
Keyword(s):  
Cardiac Muscle ◽  
Small Molecule ◽  
High Throughput ◽  
Thin Filament ◽  
High Throughput Screen ◽  
Filament Activation

scholarly journals Development of a Novel High-Throughput Screen and Identification of Small-Molecule Inhibitors of the Gα–RGS17 Protein–Protein Interaction Using AlphaScreen

2011 ◽  
Vol 16 (8) ◽  
pp. 869-877 ◽  
Author(s):  
Duncan I. Mackie ◽  
David L. Roman
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Protein Interaction ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Screening Method ◽  
Fold Increase ◽  
Rgs Proteins ◽  
High Throughput Screen ◽  
Protein Protein Interaction

In this study, the authors used AlphaScreen technology to develop a high-throughput screening method for interrogating small-molecule libraries for inhibitors of the Gαo–RGS17 interaction. RGS17 is implicated in the growth, proliferation, metastasis, and the migration of prostate and lung cancers. RGS17 is upregulated in lung and prostate tumors up to a 13-fold increase over patient-matched normal tissues. Studies show RGS17 knockdown inhibits colony formation and decreases tumorigenesis in nude mice. The screen in this study uses a measurement of the Gαo–RGS17 protein–protein interaction, with an excellent Z score exceeding 0.73, a signal-to-noise ratio >70, and a screening time of 1100 compounds per hour. The authors screened the NCI Diversity Set II and determined 35 initial hits, of which 16 were confirmed after screening against controls. The 16 compounds exhibited IC50 <10 µM in dose–response experiments. Four exhibited IC50 values <6 µM while inhibiting the Gαo–RGS17 interaction >50% when compared to a biotinylated glutathione-S-transferase control. This report describes the first high-throughput screen for RGS17 inhibitors, as well as a novel paradigm adaptable to many other RGS proteins, which are emerging as attractive drug targets for modulating G-protein-coupled receptor signaling.

scholarly journals Contributions of Ca 2+ -Independent Thin Filament Activation to Cardiac Muscle Function

2015 ◽  
Vol 109 (10) ◽  
pp. 2101-2112 ◽  
Author(s):  
Yasser Aboelkassem ◽  
Jordan A. Bonilla ◽  
Kimberly J. McCabe ◽  
Stuart G. Campbell
Keyword(s):  
Cardiac Muscle ◽  
Muscle Function ◽  
Thin Filament ◽  
Filament Activation

scholarly journals Cell‐based high‐throughput screen for small molecule inhibitors of Bax translocation

2018 ◽  
Vol 23 (3) ◽  
pp. 1784-1797 ◽  
Author(s):  
Kelvin Kai‐Wan Hui ◽  
Chesarahmia Dojo Soeandy ◽  
Stephano Chang ◽  
Frederick S. Vizeacoumar ◽  
Thomas Sun ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Small Molecule Inhibitors ◽  
High Throughput Screen

A Cell-Based, High-Throughput Screen for Small Molecule Regulators of Hepatitis C Virus Replication

2007 ◽  
Vol 132 (1) ◽  
pp. 311-320 ◽  
Author(s):  
Sun Suk Kim ◽  
Lee F. Peng ◽  
Wenyu Lin ◽  
Won-Hyeok Choe ◽  
Naoya Sakamoto ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Virus Replication ◽  
Small Molecule ◽  
High Throughput ◽  
High Throughput Screen ◽  
A Cell

scholarly journals Small Molecule Inhibitors of Phospholipase C from a Novel High-throughput Screen

2013 ◽  
Vol 288 (8) ◽  
pp. 5840-5848 ◽  
Author(s):  
Weigang Huang ◽  
Matthew Barrett ◽  
Nicole Hajicek ◽  
Stephanie Hicks ◽  
T. Kendall Harden ◽  
...  
Keyword(s):  
Phospholipase C ◽  
Small Molecule ◽  
High Throughput ◽  
Small Molecule Inhibitors ◽  
High Throughput Screen

scholarly journals The effect of altered temperature on Ca2(+)-sensitive force in permeabilized myocardium and skeletal muscle. Evidence for force dependence of thin filament activation.

1990 ◽  
Vol 96 (6) ◽  
pp. 1221-1245 ◽  
Author(s):  
N K Sweitzer ◽  
R L Moss
Keyword(s):  
Skeletal Muscle ◽  
Cardiac Muscle ◽  
Thin Filament ◽  
Calcium Sensitivity ◽  
Differential Sensitivity ◽  
Fiber Types ◽  
Tension Development ◽  
Maximum Tension ◽  
Filament Activation ◽  
Muscle Types

The effect of changes in temperature on the calcium sensitivity of tension development was examined in permeabilized cellular preparations of rat ventricle and rabbit psoas muscle. Maximum force and Ca2+ sensitivity of force development increased with temperature in both muscle types. Cardiac muscle was more sensitive to changes in temperature than skeletal muscle in the range 10-15 degrees C. It was postulated that the level of thin filament activation may be decreased by cooling. To investigate this possibility, troponin C (TnC) was partially extracted from both muscle types, thus decreasing the level of thin filament activation independent of temperature and, at least in skeletal muscle fibers, decreasing cooperative activation of the thin filament as well. TnC extraction from cardiac muscle reduced the calcium sensitivity of tension less than did extraction of TnC from skeletal muscle. In skeletal muscle the midpoint shift of the tension-pCa curve with altered temperature was greater after TnC extraction than in control fibers. Calcium sensitivity of tension development was proportional to the maximum tension generated in cardiac or skeletal muscle under all conditions studied. Based on these results, we conclude that (a) maximum tension-generating capability and calcium sensitivity of tension development are related, perhaps causally, in fast skeletal and cardiac muscles, and (b) thin filament activation is less cooperative in cardiac muscle than in skeletal muscle, which explains the differential sensitivity of the two fiber types to temperature and TnC extraction. Reducing thin filament cooperativity in skeletal muscle by TnC extraction results in a response to temperature similar to that of control cardiac cells. This study provides evidence that force levels in striated muscle influence the calcium binding affinity of TnC.

scholarly journals A Target-Based High Throughput Screen Yields Trypanosoma brucei Hexokinase Small Molecule Inhibitors with Antiparasitic Activity

2010 ◽  
Vol 4 (4) ◽  
pp. e659 ◽  
Author(s):  
Elizabeth R. Sharlow ◽  
Todd A. Lyda ◽  
Heidi C. Dodson ◽  
Gabriela Mustata ◽  
Meredith T. Morris ◽  
...  
Keyword(s):  
Trypanosoma Brucei ◽  
Small Molecule ◽  
High Throughput ◽  
Small Molecule Inhibitors ◽  
High Throughput Screen ◽  
Antiparasitic Activity

scholarly journals A high-throughput screen to identify novel small molecule inhibitors of the Werner Syndrome Helicase-Nuclease (WRN)

PLoS ONE ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. e0210525 ◽  
Author(s):  
Joshua A. Sommers ◽  
Tomasz Kulikowicz ◽  
Deborah L. Croteau ◽  
Thomas Dexheimer ◽  
Dorjbal Dorjsuren ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Small Molecule Inhibitors ◽  
Werner Syndrome ◽  
High Throughput Screen

scholarly journals Small-Molecule Inhibitors of toxT Expression in Vibrio cholerae

mBio ◽  
2013 ◽  
Vol 4 (4) ◽  
Author(s):  
Rebecca Anthouard ◽  
Victor J. DiRita
Keyword(s):  
Cholera Toxin ◽  
Vibrio Cholerae ◽  
Small Molecule ◽  
High Throughput ◽  
Small Molecule Inhibitors ◽  
Severe Disease ◽  
Virulence Gene ◽  
High Throughput Screen ◽  
Content Type ◽  
Oral Rehydration

ABSTRACTVibrio cholerae, a Gram-negative bacterium, infects humans and causes cholera, a severe disease characterized by vomiting and diarrhea. These symptoms are primarily caused by cholera toxin (CT), whose production byV. choleraeis tightly regulated by the virulence cascade. In this study, we designed and carried out a high-throughput chemical genetic screen to identify inhibitors of the virulence cascade. We identified three compounds, which we named toxtazin A and toxtazin B and Bʹ, representing two novel classes oftoxTtranscription inhibitors. All three compounds reduce production of both CT and the toxin-coregulated pilus (TCP), an important colonization factor. We present evidence that toxtazin A works at the level of thetoxTpromoter and that toxtazins B and Bʹ work at the level of thetcpPpromoter. Treatment with toxtazin B results in a 100-fold reduction in colonization in an infant mouse model of infection, though toxtazin A did not reduce colonization at the concentrations tested. These results add to the growing body of literature indicating that small-molecule inhibitors of virulence genes could be developed to treat infections, as alternatives to antibiotics become increasingly needed.IMPORTANCEV. choleraecaused more than 580,000 infections worldwide in 2011 alone (WHO, Wkly. Epidemiol. Rec. 87:289-304, 2012). Cholera is treated with an oral rehydration therapy consisting of water, glucose, and electrolytes. However, asV. choleraeis transmitted via contaminated water, treatment can be difficult for communities whose water source is contaminated. In this study, we address the need for new therapeutic approaches by targeting the production of the main virulence factor, cholera toxin (CT). The high-throughput screen presented here led to the identification of two novel classes of inhibitors of the virulence cascade inV. cholerae, toxtazin A and toxtazins B and Bʹ. We demonstrate that (i) small-molecule inhibitors of virulence gene production can be identified in a high-throughput screen, (ii) targeting virulence gene production is an effective therapeutic strategy, and (iii) small-molecule inhibitors can uncover unknown layers of gene regulation, even in well-studied regulatory cascades.

Sign in / Sign up

Export Citation Format

Share Document