scholarly journals A Quantitative Proteomics-Based Competition Binding Assay to Characterize pITAM–Protein Interactions

2013 ◽  
Vol 85 (10) ◽  
pp. 5071-5077 ◽  
Author(s):  
Lianghai Hu ◽  
Li Yang ◽  
Andrew M. Lipchik ◽  
Robert L. Geahlen ◽  
Laurie L. Parker ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Quantitative Proteomics ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

Fluorescence polarization-based competition binding assay for c-Jun N-terminal kinases 1 and 2

2017 ◽  
Vol 532 ◽  
pp. 26-28 ◽  
Author(s):  
Francesco Ansideri ◽  
Marcel Dammann ◽  
Frank M. Boeckler ◽  
Pierre Koch
Keyword(s):  
Fluorescence Polarization ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

scholarly journals Discovery of BAZ2A Bromodomain Ligands

2017 ◽  
Author(s):  
Dimitrios Spiliotopoulos ◽  
Eike-Christian Wamhoff ◽  
Graziano Lolli ◽  
Christoph Rademacher ◽  
Amedeo Caflisch
Keyword(s):  
Small Molecules ◽  
Binding Assay ◽  
Side Chain ◽  
Binding Modes ◽  
Zinc Finger Domain ◽  
Ligand Efficiency ◽  
Competition Binding Assay ◽  
Energy Evaluation ◽  
Natural Ligand ◽  
Competition Binding

The bromodomain adjacent to zinc finger domain protein 2A (BAZ2A) is implicated in aggressive prostate cancer. The BAZ2A bromodomain is a challenging target because of the shallow pocket of its natural ligand, the acetylated side chain of lysine. Here, we report the successful screening of a library of nearly 1500 small molecules by high-throughput docking and force field-based binding-energy evaluation. For seven of the 20 molecules selected in silico, evidence of binding to the BAZ2A bromodomain is provided by ligand-observed NMR spectroscopy. Two of these compounds show a favorable ligand efficiency of 0.42 kcal/mol per non-hydrogen atom in a competition-binding assay. The crystal structures of the BAZ2A bromodomain in complex with four fragment hits validate the predicted binding modes. The binding modes of compounds 1 and 3 are compatible with ligand growing for optimization of affinity for BAZ2A and selectivity against the close homologue BAZ2B.

scholarly journals Development of a Competition-Binding Assay to Determine Binding Affinity of Molecules to Neuromelanin via Fluorescence Spectroscopy

Biomolecules ◽  
2019 ◽  
Vol 9 (5) ◽  
pp. 175 ◽  
Author(s):  
Jackson Fink ◽  
Heather Pathak ◽  
John Smith ◽  
Cindy Achat-Mendes ◽  
Robert L. Haining
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Binding Affinity ◽  
Dopaminergic Neurons ◽  
Binding Assay ◽  
Pathological State ◽  
Polymeric Form ◽  
Competition Binding Assay ◽  
Competition Binding ◽  
6 Hydroxydopamine

Neuromelanin, the polymeric form of dopamine which accumulates in aging neuronal tissue, is increasingly recognized as a functional and critical component of a healthy and active adult human brain. Notorious in plant and insect literature for their ability to bind and retain amines for long periods of time, catecholamine polymers known colloquially as ‘melanins’ are nevertheless curiously absent from most textbooks regarding biochemistry, neuroscience, and evolution. Recent research has brought attention to the brain pigment due to its possible role in neurodegeneration. This linkage is best illustrated by Parkinson’s disease, which is characterized by the loss of pigmented dopaminergic neurons and the ‘white brain’ pathological state. As such, the ability to determine the binding affinity of neurotoxic agents, as well as any potential specific endogenous ligands to neuromelanin are of interest and potential value. Neuromelanin has been shown to have saturable binding interactions with nicotine as monitored by a fluorimeter. This interaction provides a signal to allow for a competition-binding assay with target molecules which do not themselves produce signal. The current report establishes the viability of this competition assay toward three compounds with central relevance to Parkinson’s disease. The Kd of binding toward neuromelanin by methyl-phenyl-pyridinium ion (MPP+), dopamine, and 6-hydroxydopamine were found to be 1 mM, 0.05 mM, and 0.1 mM, respectively in the current study. In addition, we demonstrate that 6-hydroxydopamine polymerizes to form neuromelanin granules in cultured dopaminergic neurons that treated with 2,4,5-trihydroxy-l-phenylalanine. Immunohistochemical analysis using fluor-tagged anti-dopamine antibodies suggests that the incorporation of 6-hydroxydopamine (following internalization and decarboxylation analogous to levodopa and dopamine) alters the localized distribution of bound dopamine in these cells.

A new competition binding assay for determination of the cAMP content of human leukocytes

1991 ◽  
Vol 178 (3) ◽  
pp. 980-984 ◽  
Author(s):  
Timo P.L. Zomerdijk ◽  
Peter H. Nibbering ◽  
Anja C. Bezemer ◽  
Clemens W.G.M. Löwik ◽  
Ralph van Furth
Keyword(s):  
Binding Assay ◽  
Camp Content ◽  
Human Leukocytes ◽  
Competition Binding Assay ◽  
Competition Binding

Competition Binding Assay in Cell Culture for Identification of Epitopes on Enveloped and Naked Viruses

1989 ◽  
Vol 271 (2) ◽  
pp. 237-243 ◽  
Author(s):  
Tom A.M. Oosterlaken ◽  
Fer Vlaspolder ◽  
René Fransen ◽  
Theo Harmsen ◽  
Cornelis A. Kraaijeveld ◽  
...  
Keyword(s):  
Cell Culture ◽  
Binding Assay ◽  
Competition Binding Assay ◽  
Competition Binding

Fluorescent-Labeled Selective Adenosine A2B Receptor Antagonist Enables Competition Binding Assay by Flow Cytometry

2018 ◽  
Vol 61 (10) ◽  
pp. 4301-4316 ◽  
Author(s):  
Meryem Köse ◽  
Sabrina Gollos ◽  
Tadeusz Karcz ◽  
Amelie Fiene ◽  
Fabian Heisig ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Receptor Antagonist ◽  
Binding Assay ◽  
Adenosine A2b Receptor ◽  
Competition Binding Assay ◽  
A2b Receptor ◽  
Competition Binding ◽  
Adenosine A2b
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