The Significance of G Protein-Coupled Receptor Crystallography for Drug Discovery

Salon, John; Lodowski, David T.; Palczewski, Krzysztof

doi:10.1124/pr.110.003350

Cited by 201 publications

(193 citation statements)

References 322 publications

(354 reference statements)

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“…Homology modeling is further complicated by the fact that the available crystal structures only provide snapshots of receptor conformations, and very often, the loop regions are poorly resolved. Even with the most accurate models, based on templates with >50 % homology, it remains difficult to predict the exact ligand binding site [32][33][34]. This is even more difficult if the ligand is a small, uncharged molecule, such as adenine.…”

Section: Discussionmentioning

confidence: 99%

“…Four injections were performed before serum collection. To gain better specificity, additional antibodies were raised against the following peptides: [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43] (N-term), 181-194 (extracellular loop 2 (ECL2)), 222-235 (intracellular loop 3 (ICL3)), 295-314 (C-terminal), and 313-331 (C-terminal). These antibodies were produced in two rabbits each by Thermo Scientific.…”

Section: Antibodiesmentioning

confidence: 99%

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The rat adenine receptor: pharmacological characterization and mutagenesis studies to investigate its putative ligand binding site

Knospe

Müller

Rosa

et al. 2013

Purinergic Signalling

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The rat adenine receptor (rAdeR) was the first member of a family of G protein-coupled receptors (GPCRs) activated by adenine and designated as P0-purine receptors. The present study aimed at gaining insights into structural aspects of ligand binding and function of the rAdeR. We exchanged amino acid residues predicted to be involved in ligand binding (Phe110 3.24 47 , were found to be crucial for activation since their alanine mutants did not respond to adenine. Moreover we showed that-in contrast to most other rhodopsin-like GPCRs-the rAdeR does not contain essential disulfide bonds since preincubation with dithiothreitol neither altered adenine binding in Sf9 cell membranes, nor adenineinduced inhibition of adenylate cyclase in 1321N1 astrocytoma cells transfected with the rAdeR. To detect rAdeRs by Western blot analysis, we developed a specific antibody. Finally, we were able to show that the extended N-terminal sequence of the rAdeR constitutes a putative signal peptide of unknown function that is cleaved off in the mature receptor. Our results provide important insights into this new, poorly investigated family of purinergic receptors.

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Section: Discussionmentioning

confidence: 99%

Section: Antibodiesmentioning

confidence: 99%

The rat adenine receptor: pharmacological characterization and mutagenesis studies to investigate its putative ligand binding site

Knospe

Müller

Rosa

et al. 2013

Purinergic Signalling

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show abstract

“…Amino (N)-terminal fragment is extracellular and the carboxyl (C)-terminal tail is intracellular. In the recent years this common structural topology was also confirmed by three-dimensional crystal structure of some 7TMR members (reviewed by (Salon et al, 2011)). Additionally, 7TMRs may undergo a variety of posttranslational modifications such as N-linked glycosylation, formation of disulfide bonds, palmitoylation and phosphorylation.…”

mentioning

confidence: 86%

Quantitative Assessment of Seven Transmembrane Receptors (7TMRs) Oligomerization by Bioluminescence Resonance Energy Transfer (BRET) Technology

Kubale¹,

Drinovec²,

Vrecl³

2012

Bioluminescence - Recent Advances in Oceanic Measurements and Laboratory Applications

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“…All these receptor modulation pathways, which alter localization and protein conformation, are important factors in the use of GPCRs as biomarkers. Interestingly, most of the currently available medicines, whether agonists, antagonists, inverse agonists or allosteric modulators, interact with GPCRs [10][11][12].…”

Section: Role Of Membrane Proteins In Health Disease and Medical Diagnmentioning

confidence: 99%

Cell Surface Membrane Proteins as Personalized Biomarkers: Where we Stand and Where we are Headed

et al. 2013

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Personalized medicine requires the development of a wide array of biomarker diagnostic assays, reflecting individual variations and thus allowing tailored therapeutic interventions. Membrane proteins comprise approximately 30% of total human proteins; they play a key role in various physiological functions and pathological conditions, although, currently, only a limited number of membrane proteins are applied as biomarkers. In many normal tissues, cell surface membrane proteins are not easily accessible for diagnostic sampling, and tumor-derived membrane preparations – while serving as potential tumor biomarkers – may not reflect physiological protein expression. In addition to post-translational modifications, which may include glycosylation, phosphorylation and lipid modifications, the trafficking of membrane proteins is also regulated. Moreover, a tight cellular quality control monitors membrane protein maturation, and continuous removal and reinsertion, involving special signaling systems, occurs in many cases. However, cell surface membrane proteins already serve as valuable prognostic and predicative biomarkers, for example, in hematological and immunological diseases, by the determination of the cluster of differentiation markers. In this review, we demonstrate the relevance of cell surface membrane biomarkers in various diseases and call attention to the potential application of red blood cell (erythrocyte) membrane proteins in this regard. Surprisingly, red blood cells express hundreds of membrane proteins, which seem to reflect a general genetic and regulatory background, and may serve as relatively stable and easily accessible personalized membrane biomarkers. Quantitative membrane protein detection in red blood cells by flow cytometry may bring a breakthrough in this regard.

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The Significance of G Protein-Coupled Receptor Crystallography for Drug Discovery

Cited by 201 publications

References 322 publications

The rat adenine receptor: pharmacological characterization and mutagenesis studies to investigate its putative ligand binding site

The rat adenine receptor: pharmacological characterization and mutagenesis studies to investigate its putative ligand binding site

Quantitative Assessment of Seven Transmembrane Receptors (7TMRs) Oligomerization by Bioluminescence Resonance Energy Transfer (BRET) Technology

Cell Surface Membrane Proteins as Personalized Biomarkers: Where we Stand and Where we are Headed

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