Frank Bernhard
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View article: Cryo-EM structure of a cell-free synthesized full-length human β1-adrenergic receptor in complex with Gs
Cryo-EM structure of a cell-free synthesized full-length human β1-adrenergic receptor in complex with Gs Open
The third intracellular loop (ICL3) of the β1-adrenergic receptor (β1AR) plays a critical role in regulating G protein coupling, yet the structural basis has remained unclear due to truncations of ICL3 in all available structures of the β1…
View article: Cryo-EM structure of a cell-free synthesized full-length human β1-adrenergic receptor in complex with G<sub>s</sub>
Cryo-EM structure of a cell-free synthesized full-length human β1-adrenergic receptor in complex with G<sub>s</sub> Open
SUMMARY The third intracellular loop (ICL3) of the β1-adrenergic receptor (β1AR) plays a critical role in regulating G protein coupling, yet the structural basis has remained unclear due to truncations of ICL3 in all available structures o…
View article: Flavopiridol restores granulopoiesis in experimental models of severe congenital neutropenia
Flavopiridol restores granulopoiesis in experimental models of severe congenital neutropenia Open
Severe congenital neutropenia (CN) patients require life-long treatment with recombinant human granulocyte colony-stimulating factor (rhG-CSF), but some show no response. We sought to establish a therapy for CN that targets signaling pathw…
View article: Transporter function characterization via continuous-exchange cell-free synthesis and solid supported membrane-based electrophysiology
Transporter function characterization via continuous-exchange cell-free synthesis and solid supported membrane-based electrophysiology Open
Functional characterization of transporters is impeded by the high cost and technical challenges of current transporter assays. Thus, in this work, we developed a new characterization workflow that combines cell-free protein synthesis (CFP…
View article: Cryo-EM structure of cell-free synthesized human histamine 2 receptor/Gs complex in nanodisc environment
Cryo-EM structure of cell-free synthesized human histamine 2 receptor/Gs complex in nanodisc environment Open
Here we describe the cryo-electron microscopy structure of the human histamine 2 receptor (H 2 R) in an active conformation with bound histamine and in complex with G s heterotrimeric protein at an overall resolution of 3.4 Å. The complex …
View article: Cryo-EM structure of cell-free synthesized human histamine H<sub>2</sub>receptor coupled to heterotrimeric G<sub>s</sub>protein in lipid nanodisc environment
Cryo-EM structure of cell-free synthesized human histamine H<sub>2</sub>receptor coupled to heterotrimeric G<sub>s</sub>protein in lipid nanodisc environment Open
Here we describe the cryo-electron microscopy structure of the human histamine 2 receptor (H 2 R) in an active conformation with bound histamine and in complex with G s heterotrimeric protein at an overall resolution of 3.4 Å. The complex …
View article: <i>In vitro</i> characterization of the phage lysis protein MS2-L
<i>In vitro</i> characterization of the phage lysis protein MS2-L Open
Background: The peptide MS2-L represents toxins of the ssRNA Leviviridae phage family and consists of a predicted N-terminal soluble domain followed by a transmembrane domain. MS2-L mediates bacterial cell lysis through the formation of la…
View article: E. coli “Stablelabel” S30 lysate for optimized cell-free NMR sample preparation
E. coli “Stablelabel” S30 lysate for optimized cell-free NMR sample preparation Open
Cell-free (CF) synthesis with highly productive E. coli lysates is a convenient method to produce labeled proteins for NMR studies. Despite reduced metabolic activity in CF lysates, a certain scrambling of supplied isotope labels is still …
View article: Electroneutral Polymer Nanodiscs Enable Interference‐Free Probing of Membrane Proteins in a Lipid‐Bilayer Environment
Electroneutral Polymer Nanodiscs Enable Interference‐Free Probing of Membrane Proteins in a Lipid‐Bilayer Environment Open
Membrane proteins can be examined in near‐native lipid‐bilayer environments with the advent of polymer‐encapsulated nanodiscs. These nanodiscs self‐assemble directly from cellular membranes, allowing in vitro probing of membrane proteins w…
View article: Transfer mechanism of cell-free synthesized membrane proteins into mammalian cells
Transfer mechanism of cell-free synthesized membrane proteins into mammalian cells Open
Nanodiscs are emerging to serve as transfer vectors for the insertion of recombinant membrane proteins into membranes of living cells. In combination with cell-free expression technologies, this novel process opens new perspectives to anal…
View article: Membrane insertion mechanism and molecular assembly of the bacteriophage lysis toxin ΦX174‐E
Membrane insertion mechanism and molecular assembly of the bacteriophage lysis toxin ΦX174‐E Open
The bacteriophage ΦX174 causes large pore formation in Escherichia coli and related bacteria. Lysis is mediated by the small membrane‐bound toxin ΦX174‐E, which is composed of a transmembrane domain and a soluble domain. The toxin requires…
View article: Inside Cover: The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes (Angew. Chem. Int. Ed. 52/2020)
Inside Cover: The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes (Angew. Chem. Int. Ed. 52/2020) Open
NMR spectroscopy sheds light on receptor activation: Molecular switches are conserved motifs of G protein-coupled receptors that undergo structural changes during activation. Tryptophan residues are often part of these molecular switches a…
View article: Integration of Cell-Free Expression and Solid-State NMR to Investigate the Dynamic Properties of Different Sites of the Growth Hormone Secretagogue Receptor
Integration of Cell-Free Expression and Solid-State NMR to Investigate the Dynamic Properties of Different Sites of the Growth Hormone Secretagogue Receptor Open
Cell-free expression represents an attractive method to produce large quantities of selectively labeled protein for NMR applications. Here, cell-free expression was used to label specific regions of the growth hormone secretagogue receptor…
View article: The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes
The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes Open
Dynamic structural transitions within the seven‐transmembrane bundle represent the mechanism by which G‐protein‐coupled receptors convert an extracellular chemical signal into an intracellular biological function. Here, the conformational …
View article: Rat Organic Cation Transporter 1 Contains Three Binding Sites for Substrate 1-Methyl-4-phenylpyridinium per Monomer
Rat Organic Cation Transporter 1 Contains Three Binding Sites for Substrate 1-Methyl-4-phenylpyridinium per Monomer Open
Organic cation transporters OCT1 (SLC22A1) and OCT2 (SLC22A2) are critically involved in absorption and excretion of diverse cationic drugs. Because drug-drug interactions at these transporters may induce adverse drug effects in patients, …
View article: LILBID and nESI: Different Native Mass Spectrometry Techniques as Tools in Structural Biology
LILBID and nESI: Different Native Mass Spectrometry Techniques as Tools in Structural Biology Open
Native mass spectrometry is applied for the investigation of proteins and protein complexes worldwide. The challenge in native mass spectrometry is maintaining the features of the proteins of interest, such as oligomeric state, bound ligan…
View article: The soluble loop BC region guides, but not dictates, the assembly of the transmembrane cytochrome b6
The soluble loop BC region guides, but not dictates, the assembly of the transmembrane cytochrome b6 Open
Studying folding and assembly of naturally occurring α-helical transmembrane proteins can inspire the design of membrane proteins with defined functions. Thus far, most studies have focused on the role of membrane-integrated protein region…
View article: Integral Membrane Proteins Can Be Crystallized Directly from Nanodiscs
Integral Membrane Proteins Can Be Crystallized Directly from Nanodiscs Open
Membrane-like nanodiscs (ND) have become an important tool for the cell-free expression, solubilization, folding, and in vitro structural and functional studies of membrane proteins (MPs). Direct crystallization of MPs embedded in NDs woul…
View article: Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology
Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology Open
Membrane proteins frequently assemble into higher order homo- or hetero-oligomers within their natural lipid environment. This complex formation can modulate their folding, activity as well as substrate selectivity. Non-disruptive methods …
View article: Author response: Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology
Author response: Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology Open
Article Figures and data Abstract Introduction Results Discussion Materials and methods References Decision letter Author response Article and author information Metrics Abstract Membrane proteins frequently assemble into higher order homo…
View article: The synaptic vesicle protein <scp>SV</scp>31 assembles into a dimer and transports Zn<sup>2+</sup>
The synaptic vesicle protein <span>SV</span>31 assembles into a dimer and transports Zn<sup>2+</sup> Open
The integral synaptic vesicle protein SV 31 has been shown to bind divalent cations. Here, we demonstrate that SV 31 protein synthesized within a cell‐free system binds Zn 2+ and to a lower extent Ni 2+ and Cu 2+ ions. Expression with Zn 2…