Reevaluation of fenpropimorph as a σ receptor ligand: Structure-affinity relationship studies at human σ1 receptors

Sguazzini, Elena; Schmidt, Hayden R.; Iyer, Kavita A.; Kruse, Andrew C.; Dukat, Małgorzata

doi:10.1016/j.bmcl.2017.04.088

Cited by 9 publications

(10 citation statements)

References 17 publications

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“…Membranes were prepared as described previously 52 , using a protocol adapted from that of Vilner et al 53 . In brief, Sf9 cells expressing σ 1 receptor were harvested by centrifugation and lysed by osmotic shock in a buffer containing 20 mM HEPES pH 7.5, 2 mM MgCl2,1:200,000 (v/v) benzonase nuclease (Sigma Aldrich), and cOmplete Mini EDTA-free protease inhibitor tablets (Sigma Aldrich).…”

Section: Methodsmentioning

confidence: 99%

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Structural basis for σ1 receptor ligand recognition

Schmidt

Betz

Dror

et al. 2018

Nat Struct Mol Biol

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The σ 1 receptor is a poorly understood membrane protein expressed throughout the human body. Ligands targeting the σ 1 receptor are in clinical trials for treatment of Alzheimer’s disease, ischemic stroke, and neuropathic pain. Despite this, relatively little is known regarding the σ 1 receptor’s molecular function. Here, we present crystal structures of human σ 1 receptor bound to the antagonists haloperidol and NE-100, and the agonist (+)-pentazocine, at crystallographic resolutions of 3.1 Å, 2.9 Å, and 3.1 Å respectively. These structures reveal a unique binding pose for the agonist. The structures and accompanying molecular dynamics (MD) simulations identify agonist-induced structural rearrangements in the receptor. Additionally, we show that ligand binding to σ 1 is a multistep process, rate-limited by receptor conformational change. We use MD simulations to reconstruct a ligand binding pathway involving two major conformational changes. These data provide a framework for understanding the molecular basis for σ 1 agonism.

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Section: Methodsmentioning

confidence: 99%

“…Saturation binding was performed as described previously 52 , using a method similar to that of Chu and Ruoho 54 . Briefly, membrane samples from Sf9 cells expressing wild-type or mutant σ 1 receptor prepared as described above were thawed, homogenized with a syringe, and diluted in 50 mM Tris pH 8.0.…”

Section: Methodsmentioning

confidence: 99%

Structural basis for σ1 receptor ligand recognition

Schmidt

Betz

Dror

et al. 2018

Nat Struct Mol Biol

Self Cite

118

161

View full text Add to dashboard Cite

show abstract

“…Membranes were prepared as described previously 45 , using a protocol adapted from that of Vilner et al 46 . In brief, Sf9 cells expressing σ 1 receptor were harvested by centrifugation and lysed by osmotic shock in a buffer containing 20 mM HEPES pH 7.5, 2 mM MgCl2,1:200,000 (v/v) benzonase nuclease (Sigma Aldrich), and cOmplete Mini EDTA-free protease inhibitor tablets (Sigma Aldrich).…”

Section: Preparation Of Membranes For Radioligand Bindingmentioning

confidence: 99%

“…Saturation binding was performed as described previously 45 , using a method similar to that of Chu and Ruoho 47 . Briefly, membrane samples from Sf9 cells expressing wild-type or mutant σ 1 receptor prepared as described above were thawed, homogenized with a syringe, and diluted in 50 mM Tris pH 8.0.…”

Section: Saturation Binding In Sf9 Membranesmentioning

confidence: 99%

Structural basis for σ₁ receptor ligand recognition

Schmidt

Betz

Dror

et al. 2018

Preprint

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The σ 1 receptor is a poorly understood integral membrane protein expressed in most cells and tissues in the human body. It has been shown to modulate the activity of other membrane proteins such as ion channels and G protein-coupled receptors 1-4 , and ligands targeting the σ 1 receptor are currently in clinical trials for treatment of Alzheimer's disease 5 , ischemic stroke 6 , and neuropathic pain 7 . Despite its importance, relatively little is known regarding σ 1 receptor function at the molecular level. Here, we present crystal structures of the human σ 1 receptor bound to the classical antagonists haloperidol and NE-100, as well as the agonist (+)-pentazocine, at crystallographic resolutions of 3.1 Å, 2.9 Å, and 3.1 Å respectively. These structures reveal a unique binding pose for the agonist. The structures and accompanying molecular dynamics (MD) simulations demonstrate that the agonist induces subtle structural rearrangements in the receptor. In addition, we show that ligand binding and dissociation from σ 1 is a multistep process, with extraordinarily slow kinetics limited by receptor conformational change. We use MD simulations to reconstruct a ligand binding pathway that requires two major conformational changes. Taken together, these data provide a framework for understanding the molecular basis for agonist action at σ 1 .Discovered in 1976 8 , the σ 1 receptor has attracted interest because it binds a host of structurally dissimilar pharmacologically active compounds with high affinity (Fig. 1a). These include benzomorphans, antipsychotics, psychosis-inducing drugs, the antifungal agent fenpropimorph, sterols such as progesterone, and numerous other compounds 9 . These molecules contain few shared features, although most include a basic nitrogen atom flanked on two sides by longer hydrophobic moieties (typically

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“…Four ligands, R- and S -PTZ plus R - and S -PHZ, confirm the experimental observations about the importance of the ligand's stereochemistry. According to Sguazzine et al [ 47 ], the stereochemistry of the fenpropimorph is not important for the binding to the σ 1 receptor, while the results found by Rossi et al [ 29 ] suggest that in the case of the ( R , S )-RC-33 ligands, there are preferences toward the S -conformation when the OH group is present in the chirality center. Hence, it is interesting how the PTZ and PHZ chirality affects the ligand–protein interactions.…”

Section: Resultsmentioning

confidence: 99%

Structural Insights into σ1 Receptor Interactions with Opioid Ligands by Molecular Dynamics Simulations

et al. 2018

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Despite considerable advances over the past years in understanding the mechanisms of action and the role of the σ1 receptor, several questions regarding this receptor remain unanswered. This receptor has been identified as a useful target for the treatment of a diverse range of diseases, from various central nervous system disorders to cancer. The recently solved issue of the crystal structure of the σ1 receptor has made elucidating the structure–activity relationship feasible. The interaction of seven representative opioid ligands with the crystal structure of the σ1 receptor (PDB ID: 5HK1) was simulated for the first time using molecular dynamics (MD). Analysis of the MD trajectories has provided the receptor–ligand interaction fingerprints, combining information on the crucial receptor residues and frequency of the residue–ligand contacts. The contact frequencies and the contact maps suggest that for all studied ligands, the hydrophilic (hydrogen bonding) interactions with Glu172 are an important factor for the ligands’ affinities toward the σ1 receptor. However, the hydrophobic interactions with Tyr120, Val162, Leu105, and Ile124 also significantly contribute to the ligand–receptor interplay and, in particular, differentiate the action of the agonistic morphine from the antagonistic haloperidol.

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Reevaluation of fenpropimorph as a σ receptor ligand: Structure-affinity relationship studies at human σ1 receptors

Cited by 9 publications

References 17 publications

Structural basis for σ1 receptor ligand recognition

Structural basis for σ1 receptor ligand recognition

Structural basis for σ₁ receptor ligand recognition

Structural Insights into σ1 Receptor Interactions with Opioid Ligands by Molecular Dynamics Simulations

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