Functional GPCR microarrays.

Yulong Hong; Brian L Webb; Hui Su; Eric J Mozdy; Ye Fang; Qi Wu; Li Liu; Jonathan Beck; Ann M Ferrie; Srikanth Raghavan; John Mauro; Alain Carre; Dirk Müeller; Fang Lai; Brian Rasnow; Michael Johnson; Hosung Min; John Salon; Joydeep Lahiri

doi:10.1021/ja055101h

PubMed · 16262381

Functional GPCR microarrays.

Yulong Hong·Brian L Webb·Hui Su·Eric J Mozdy·Ye Fang·Qi Wu·Li Liu·Jonathan Beck·Ann M Ferrie·Srikanth Raghavan·John Mauro·Alain Carre·Dirk Müeller·Fang Lai·Brian Rasnow·Michael Johnson·Hosung Min·John Salon·Joydeep Lahiri

View original source ↗Export metadata ↓

Abstract

This paper describes G-protein-coupled receptor (GPCR) microarrays on porous glass substrates and functional assays based on the binding of a europium-labeled GTP analogue. The porous glass slides were made by casting a glass frit on impermeable glass slides and then coating with gamma-aminopropyl silane (GAPS). The emitted fluorescence was captured on an imager with a time-gated intensified CCD detector. Microarrays of the neurotensin receptor 1, the cholinergic receptor muscarinic 2, the opioid receptor mu, and the cannabinoid receptor 1 were fabricated by pin printing. The selective agonism of each of the receptors was observed. The screening of potential antagonists was demonstrated using a cocktail of agonists. The amount of activation observed was sufficient to permit determinations of EC50 and IC50. Such microarrays could potentially streamline drug discovery by helping integrate primary screening with selectivity and safety screening without compromising the essential functional information obtainable from cellular assays.

Explore related subjects

Drug Evaluation, Preclinical Fluorescent Dyes Ligands Protein Array Analysis Receptors, G-Protein-Coupled Journal Article

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Yulong Hong, Brian L Webb, Hui Su, Eric J Mozdy, Ye Fang, Qi Wu, Li Liu, Jonathan Beck, Ann M Ferrie, Srikanth Raghavan, John Mauro, Alain Carre, Dirk Müeller, Fang Lai, Brian Rasnow, Michael Johnson, Hosung Min, John Salon, Joydeep Lahiri. 2005-11-09. Functional GPCR microarrays.. https://doi.org/10.1021/ja055101h

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2006

Phosphonic acid-containing analogues of mycophenolic acid as inhibitors of IMPDH.

The design, synthesis, and IMPDH inhibitory activity of a series of phosphonic acid-containing analogues of mycophenolic acid are described.

Drug Evaluation, Preclinical↗

PubMed2006

Research strategies for safety evaluation of nanomaterials. Part VI. Characterization of nanoscale particles for toxicological evaluation.

To properly assign mechanisms or causes for toxic effects of nanoscale materials, their properties and characteristics both outside and within the biological environment must be well understood. Scientists have many tools for studying the size, shape, and surface properties of particulates outside of the physiological environment; however, it is difficult to measure many of these same properties in situ without perturbing the environment, leading to spurious findings. Characterizing nanoparticle systems in situ can be further complicated by an organism's active clearance, defense, and/or immune responses. As toxicologists begin to examine nanomaterials in a systematic fashion, there is consensus that a series of guidelines or recommended practices is necessary for basic characterization of nanomaterials. These recommended practices should be developed jointly by physical scientists skilled in nano characterization and biological scientists experienced in toxicology research. In this article, basic nanoparticle characterization techniques are discussed, along with the some of the issues and implications associated with measuring nanoparticle properties and their interactions with biological systems. Recommendations regarding how best to approach nanomaterial characterization include using proper sampling and measurement techniques, forming multidisciplinary teams, and making measurements as close to the biological action point as possible.

Drug Evaluation, Preclinical↗

PubMed2006

Identification by 19F NMR of traditional Chinese medicinal plants possessing prolyl oligopeptidase inhibitory activity.

Prolyl oligopeptidase is a cytosolic serine peptidase that hydrolyzes proline-containing peptides at the carboxy termini of the proline residues. This peptidase has been associated with schizophrenia, bipolar affective disorder, and related neuropsychiatric disorders and might therefore have important clinical implications. Traditional Chinese medicinal (TCM) plants provide a rich source of unexplored compounds for strategies to find novel POP inhibitors, but the traditional methodologies used to identify POP inhibitors could have some limitations when working with natural products: interference with the colorimetric or fluorimetric detection methods commonly used to screen for POP inhibitors can result in the generation of false positives or false negatives. Since NMR screening is less prone to such interference, we decided to explore the use of 19F NMR to screen for POP inhibitors. We synthesized a new 19F-labeled POP substrate--Z-Gly-Pro-Phe-4(CF3)-NH2--and used it to search for new POP inhibitors in TCM plant extracts. We identified several plants with high POP-inhibitory activity and show here that the combination of 19F NMR and TCM plant extracts is a useful tool for identifying new POP inhibitors.

Drug Evaluation, Preclinical↗