22 results on '"Stix R"'
Search Results
2. DHHC20 palmitoyl-transferase reshapes the membrane to foster catalysis
- Author
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Stix, R., primary, Song, J., additional, Banerjee, A., additional, and Faraldo-Gómez, J.D., additional
- Published
- 2019
- Full Text
- View/download PDF
3. Das Arbeitsdiagramm des läufergespeisten Drehstrom-Nebenschlußmotors
- Author
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Stix, R.
- Published
- 1939
- Full Text
- View/download PDF
4. Zusätzliche Kommutierungsverluste in Gleichstromwicklungen bei endlicher Stromwendedauer
- Author
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Stix, R.
- Published
- 1930
- Full Text
- View/download PDF
5. A critical appraisal of methods to determine failure probabilities
- Author
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Schuëller, G.I., primary and Stix, R., additional
- Published
- 1987
- Full Text
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6. Relation of “correct” weight and blood findings to physicans' estimates of nutrition of school children
- Author
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STIX, R
- Published
- 1934
- Full Text
- View/download PDF
7. Distinct mechanisms of inhibition of Kv2 potassium channels by tetraethylammonium and RY785.
- Author
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Zhang S, Stix R, Orabi EA, Bernhardt N, and Faraldo-Gomez JD
- Abstract
Voltage-gated K+ channels play central roles in human physiology, both in health and disease. A repertoire of inhibitors that are both potent and specific would therefore be of great value, not only as pharmacological agents but also as research tools. The small molecule RY785 has been described as particularly promising in this regard, as it selectively inhibits channels in the Kv2 subfamily with high potency. Kv2 channels are expressed in multiple cell types in humans, and are of particular importance for neuronal function. The mechanism of action of RY785 has not yet been determined at the molecular level, but functional studies indicate it differs from that of less specific inhibitors, such as quaternary-ammonium compounds or aminopyridines; RY785 is distinct also in that it is electroneutral. To examine this mechanism at the single-molecule level, we have carried out a series of all-atom molecular dynamics simulations based on the experimental structure of the Kv2.1 channel in the activated, open state. First, we report a 25-microsecond trajectory calculated in the absence of any inhibitor, under an applied voltage of 100 mV, which demonstrates outward K+ flow under simulation conditions at rates comparable to experimental measurements. Additional simulations in which either RY785 or tetraethylammonium (TEA) is introduced in solution show both inhibitors spontaneously enter the channel through the cytoplasmic gate, with distinct effects. In agreement with prior structural studies, we observe that TEA binds to a site adjacent to the selectivity filter, on the pore axis, thereby blocking the flow of K+ ions. RY785, by contrast, binds to the channel walls, off-axis, and allows K+ flow while the cytoplasmic gate remains open. The observed mode of RY785 binding, however, indicates that its mechanism of action is to stabilize and occlude a semi-open state of the gate, by bridging hydrophobic protein-protein interactions therein; this hypothesis would explain the puzzling experimental observation that RY785 recognition influences the gating currents generated by the voltage sensors, 3 nm away.
- Published
- 2024
- Full Text
- View/download PDF
8. Eukaryotic Kv channel Shaker inactivates through selectivity filter dilation rather than collapse.
- Author
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Stix R, Tan XF, Bae C, Fernández-Mariño AI, Swartz KJ, and Faraldo-Gómez JD
- Subjects
- Cryoelectron Microscopy, Dilatation, Ion Channel Gating physiology, Potassium Channels, Voltage-Gated
- Abstract
Eukaryotic voltage-gated K
+ channels have been extensively studied, but the structural bases for some of their most salient functional features remain to be established. C-type inactivation, for example, is an auto-inhibitory mechanism that confers temporal resolution to their signal-firing activity. In a recent breakthrough, studies of a mutant of Shaker that is prone to inactivate indicated that this process entails a dilation of the selectivity filter, the narrowest part of the ion conduction pathway. Here, we report an atomic-resolution cryo-electron microscopy structure that demonstrates that the wild-type channel can also adopt this dilated state. All-atom simulations corroborate this conformation is congruent with the electrophysiological characteristics of the C-type inactivated state, namely, residual K+ conductance and altered ion specificity, and help rationalize why inactivation is accelerated or impeded by certain mutations. In summary, this study establishes the molecular basis for an important self-regulatory mechanism in eukaryotic K+ channels, laying a solid foundation for further studies.- Published
- 2023
- Full Text
- View/download PDF
9. Mechanism of 4-aminopyridine inhibition of the lysosomal channel TMEM175.
- Author
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Oh S, Stix R, Zhou W, Faraldo-Gómez JD, and Hite RK
- Subjects
- Humans, Cryoelectron Microscopy, Lysosomes metabolism, Water metabolism, 4-Aminopyridine pharmacology, Potassium Channels metabolism
- Abstract
Transmembrane protein 175 (TMEM175) is an evolutionarily distinct lysosomal cation channel whose mutation is associated with the development of Parkinson's disease. Here, we present a cryoelectron microscopy structure and molecular simulations of TMEM175 bound to 4-aminopyridine (4-AP), the only known small-molecule inhibitor of TMEM175 and a broad K
+ channel inhibitor, as well as a drug approved by the Food and Drug Administration against multiple sclerosis. The structure shows that 4-AP, whose mode of action had not been previously visualized, binds near the center of the ion conduction pathway, in the open state of the channel. Molecular dynamics simulations reveal that this binding site is near the middle of the transmembrane potential gradient, providing a rationale for the voltage-dependent dissociation of 4-AP from TMEM175. Interestingly, bound 4-AP rapidly switches between three predominant binding poses, stabilized by alternate interaction patterns dictated by the twofold symmetry of the channel. Despite this highly dynamic binding mode, bound 4-AP prevents not only ion permeation but also water flow. Together, these studies provide a framework for the rational design of novel small-molecule inhibitors of TMEM175 that might reveal the role of this channel in human lysosomal physiology both in health and disease.- Published
- 2022
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10. Structure of the Shaker Kv channel and mechanism of slow C-type inactivation.
- Author
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Tan XF, Bae C, Stix R, Fernández-Mariño AI, Huffer K, Chang TH, Jiang J, Faraldo-Gómez JD, and Swartz KJ
- Abstract
Voltage-activated potassium (Kv) channels open upon membrane depolarization and proceed to spontaneously inactivate. Inactivation controls neuronal firing rates and serves as a form of short-term memory and is implicated in various human neurological disorders. Here, we use high-resolution cryo-electron microscopy and computer simulations to determine one of the molecular mechanisms underlying this physiologically crucial process. Structures of the activated Shaker Kv channel and of its W434F mutant in lipid bilayers demonstrate that C-type inactivation entails the dilation of the ion selectivity filter and the repositioning of neighboring residues known to be functionally critical. Microsecond-scale molecular dynamics trajectories confirm that these changes inhibit rapid ion permeation through the channel. This long-sought breakthrough establishes how eukaryotic K
+ channels self-regulate their functional state through the plasticity of their selectivity filters.- Published
- 2022
- Full Text
- View/download PDF
11. Bivalent recognition of fatty acyl-CoA by a human integral membrane palmitoyltransferase.
- Author
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Lee CJ, Stix R, Rana MS, Shikwana F, Murphy RE, Ghirlando R, Faraldo-Gómez JD, and Banerjee A
- Subjects
- Acyltransferases genetics, Catalytic Domain, Cell Membrane enzymology, Gene Expression Regulation, Enzymologic, Humans, Lipoylation, Models, Molecular, Molecular Dynamics Simulation, Mutation, Protein Binding, Protein Conformation, Protein Domains, Acyl Coenzyme A chemistry, Acyl Coenzyme A metabolism, Acyltransferases metabolism
- Abstract
S-acylation, also known as palmitoylation, is the most abundant form of protein lipidation in humans. This reversible posttranslational modification, which targets thousands of proteins, is catalyzed by 23 members of the DHHC family of integral membrane enzymes. DHHC enzymes use fatty acyl-CoA as the ubiquitous fatty acyl donor and become autoacylated at a catalytic cysteine; this intermediate subsequently transfers the fatty acyl group to a cysteine in the target protein. Protein S-acylation intersects with almost all areas of human physiology, and several DHHC enzymes are considered as possible therapeutic targets against diseases such as cancer. These efforts would greatly benefit from a detailed understanding of the molecular basis for this crucial enzymatic reaction. Here, we combine X-ray crystallography with all-atom molecular dynamics simulations to elucidate the structure of the precatalytic complex of human DHHC20 in complex with palmitoyl CoA. The resulting structure reveals that the fatty acyl chain inserts into a hydrophobic pocket within the transmembrane spanning region of the protein, whereas the CoA headgroup is recognized by the cytosolic domain through polar and ionic interactions. Biochemical experiments corroborate the predictions from our structural model. We show, using both computational and experimental analyses, that palmitoyl CoA acts as a bivalent ligand where the interaction of the DHHC enzyme with both the fatty acyl chain and the CoA headgroup is important for catalytic chemistry to proceed. This bivalency explains how, in the presence of high concentrations of free CoA under physiological conditions, DHHC enzymes can efficiently use palmitoyl CoA as a substrate for autoacylation., Competing Interests: The authors declare no competing interest., (Copyright © 2022 the Author(s). Published by PNAS.)
- Published
- 2022
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- View/download PDF
12. An Unbound Proline-Rich Signaling Peptide Frequently Samples Cis Conformations in Gaussian Accelerated Molecular Dynamics Simulations.
- Author
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Alcantara J, Stix R, Huang K, Connor A, East R, Jaramillo-Martinez V, Stollar EJ, and Ball KA
- Abstract
Disordered proline-rich motifs are common across the proteomes of many species and are often involved in protein-protein interactions. Proline is a unique amino acid due to the covalent bond between the backbone nitrogen and the proline side chain. The resulting five-membered ring allows proline to sample the cis state about its peptide bond, which other residues cannot do as readily. Because proline-rich disordered sequences exist as ensembles that likely include structures with the proline peptide bond in cis , a robust methodology to accurately account for these conformations in the overall ensemble is crucial. Observing the cis conformations of proline in a disordered sequence is challenging both experimentally and computationally. Nitrogen-hydrogen NMR spectroscopy cannot directly observe proline residues, which lack an amide bond, and computational methods struggle to overcome the large kinetic barrier between the cis and trans states, since isomerization usually occurs on the order of seconds. In the current work, Gaussian accelerated molecular dynamics was used to overcome this free energy barrier and simulate proline isomerization in a tetrapeptide (KPTP) and in the 12-residue proline-rich SH3 binding peptide, ArkA. We found that Gaussian accelerated molecular dynamics, when combined with a lowered peptide bond dihedral angle potential energy barrier (15 kcal/mol), allowed sufficient sampling of the proline cis and trans states on a microsecond timescale. All ArkA prolines spend a significant fraction of time in cis , leading to a more compact ensemble with less polyproline II helix structure than an ArkA ensemble with all peptide bonds in trans . The ensemble containing cis prolines also matches more closely to in vitro circular dichroism data than the all- trans ensemble. The ability of the ArkA prolines to isomerize likely affects the peptide's ability to bind its partner SH3 domain, and should be studied further. This is the first molecular dynamics simulation study of proline isomerization in a biologically relevant proline-rich sequence that we know of, and a similar protocol could be applied to study multi-proline isomerization in other proline-containing proteins to improve conformational diversity and agreement with in vitro data., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Alcantara, Stix, Huang, Connor, East, Jaramillo-Martinez, Stollar and Ball.)
- Published
- 2021
- Full Text
- View/download PDF
13. Conserved binding site in the N-lobe of prokaryotic MATE transporters suggests a role for Na + in ion-coupled drug efflux.
- Author
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Castellano S, Claxton DP, Ficici E, Kusakizako T, Stix R, Zhou W, Nureki O, Mchaourab HS, and Faraldo-Gómez JD
- Abstract
In both prokaryotes and eukaryotes, multidrug and toxic-compound extrusion (MATE) transporters catalyze the efflux of a broad range of cytotoxic compounds, including human-made antibiotics and anticancer drugs. MATEs are secondary-active antiporters, i.e. their drug-efflux activity is coupled to, and powered by, the uptake of ions down a pre-existing transmembrane electrochemical gradient. Key aspects of this mechanism, however, remain to be delineated, such as its ion specificity and stoichiometry. We previously revealed the existence of a Na+-binding site in a MATE transporter from Pyroccocus furiosus (PfMATE) and hypothesized that this site might be broadly conserved among prokaryotic MATEs. Here, we evaluate this hypothesis by analyzing VcmN and ClbM, which along with PfMATE are the only three prokaryotic MATEs whose molecular structures have been determined at resolutions better than 3 Å. Analysis of available crystallographic data and molecular dynamics simulations indeed reveal an occupied Na+-binding site in the N-terminal lobe of both structures, analogous to that identified in PfMATE. We likewise find this site to be strongly selective against K+, suggesting it is mechanistically significant. Consistent with these computational results, DEER spectroscopy measurements for multiple doubly-spin-labeled VcmN constructs demonstrate Na+-dependent changes in protein conformation. The existence of this binding site in three MATE orthologs implicates Na+ in the ion-coupled drug-efflux mechanisms of this class of transporters. These results also imply that observations of H+-dependent activity stem either from a site elsewhere in the structure, or from H+ displacing Na+ under certain laboratory conditions, as has been noted for other Na+-driven transport systems., (Published under license by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2021
- Full Text
- View/download PDF
14. A disordered encounter complex is central to the yeast Abp1p SH3 domain binding pathway.
- Author
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Gerlach GJ, Carrock R, Stix R, Stollar EJ, and Ball KA
- Subjects
- Molecular Dynamics Simulation, Protein Binding, Protein Conformation, Intrinsically Disordered Proteins chemistry, Intrinsically Disordered Proteins genetics, Intrinsically Disordered Proteins metabolism, Microfilament Proteins chemistry, Microfilament Proteins genetics, Microfilament Proteins metabolism, Saccharomyces cerevisiae Proteins chemistry, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism, src Homology Domains genetics, src Homology Domains physiology
- Abstract
Protein-protein interactions are involved in a wide range of cellular processes. These interactions often involve intrinsically disordered proteins (IDPs) and protein binding domains. However, the details of IDP binding pathways are hard to characterize using experimental approaches, which can rarely capture intermediate states present at low populations. SH3 domains are common protein interaction domains that typically bind proline-rich disordered segments and are involved in cell signaling, regulation, and assembly. We hypothesized, given the flexibility of SH3 binding peptides, that their binding pathways include multiple steps important for function. Molecular dynamics simulations were used to characterize the steps of binding between the yeast Abp1p SH3 domain (AbpSH3) and a proline-rich IDP, ArkA. Before binding, the N-terminal segment 1 of ArkA is pre-structured and adopts a polyproline II helix, while segment 2 of ArkA (C-terminal) adopts a 310 helix, but is far less structured than segment 1. As segment 2 interacts with AbpSH3, it becomes more structured, but retains flexibility even in the fully engaged state. Binding simulations reveal that ArkA enters a flexible encounter complex before forming the fully engaged bound complex. In the encounter complex, transient nonspecific hydrophobic and long-range electrostatic contacts form between ArkA and the binding surface of SH3. The encounter complex ensemble includes conformations with segment 1 in both the forward and reverse orientation, suggesting that segment 2 may play a role in stabilizing the correct binding orientation. While the encounter complex forms quickly, the slow step of binding is the transition from the disordered encounter ensemble to the fully engaged state. In this transition, ArkA makes specific contacts with AbpSH3 and buries more hydrophobic surface. Simulating the binding between ApbSH3 and ArkA provides insight into the role of encounter complex intermediates and nonnative hydrophobic interactions for other SH3 domains and IDPs in general., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2020
- Full Text
- View/download PDF
15. Structure and Mechanism of DHHC Protein Acyltransferases.
- Author
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Stix R, Lee CJ, Faraldo-Gómez JD, and Banerjee A
- Subjects
- Acylation, Crystallography, X-Ray, Humans, Models, Molecular, Protein Domains, Protein Processing, Post-Translational, Acyltransferases chemistry, Acyltransferases metabolism
- Abstract
S-acylation, whereby a fatty acid chain is covalently linked to a cysteine residue by a thioester linkage, is the most prevalent kind of lipid modification of proteins. Thousands of proteins are targets of this post-translational modification, which is catalyzed by a family of eukaryotic integral membrane enzymes known as DHHC protein acyltransferases (DHHC-PATs). Our knowledge of the repertoire of S-acylated proteins has been rapidly expanding owing to development of the chemoproteomic techniques. There has also been an increasing number of reports in the literature documenting the importance of S-acylation in human physiology and disease. Recently, the first atomic structures of two different DHHC-PATs were determined using X-ray crystallography. This review will focus on the insights gained into the molecular mechanism of DHHC-PATs from these structures and highlight representative data from the biochemical literature that they help explain., (Published by Elsevier Ltd.)
- Published
- 2020
- Full Text
- View/download PDF
16. DHHC20 Palmitoyl-Transferase Reshapes the Membrane to Foster Catalysis.
- Author
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Stix R, Song J, Banerjee A, and Faraldo-Gómez JD
- Subjects
- Acyl Coenzyme A metabolism, Acylation, Catalysis, Humans, Acyltransferases chemistry, Lipoylation
- Abstract
Cysteine palmitoylation, a form of S-acylation, is a key posttranslational modification in cellular signaling. This type of reversible lipidation occurs in both plasma and organellar membranes, and is catalyzed by a family of integral membrane proteins known as DHHC acyltransferases. The first step in the S-acylation process is the recognition of free acyl coenzyme A (acyl-CoA) from the lipid bilayer. The DHHC enzyme then becomes autoacylated at a site defined by a conserved Asp-His-His-Cys motif. This reaction entails ionization of the catalytic Cys. Intriguingly, in known DHHC structures, this catalytic Cys appears to be exposed to the hydrophobic interior of the lipid membrane, which would be highly unfavorable for a negatively charged nucleophile, thus hindering autoacylation. Here, we use biochemical and computational methods to reconcile these seemingly contradictory facts. First, we experimentally demonstrate that human DHHC20 is active when reconstituted in POPC nanodiscs. Microsecond-long all-atom molecular dynamics simulations are then calculated for human DHHC20 and for different acyl-CoA forms, also in a POPC membrane. Strikingly, we observe that human DHHC20 induces a drastic deformation in the membrane, particularly on the cytoplasmic side, where autoacylation occurs. As a result, the catalytic Cys becomes hydrated and optimally positioned to encounter the cleavage site in acyl-CoA. In summary, we hypothesize that DHHC enzymes locally reshape the membrane to foster a morphology that is specifically adapted for acyl-CoA recognition and autoacylation., (Published by Elsevier Inc.)
- Published
- 2020
- Full Text
- View/download PDF
17. Cimetidine and psoriatic arthritis.
- Author
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Manger BF, Stix R, Beck AL, and Kalden JR
- Subjects
- Humans, Arthritis drug therapy, Cimetidine therapeutic use, Guanidines therapeutic use, Psoriasis drug therapy
- Published
- 1983
18. [Effect of histamine-blocking substances on mitogen-induced lympho proliferation in rheumatic diseases].
- Author
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Manger BJ, Stix R, Platzer E, Hess B, and Kalden JR
- Subjects
- Dose-Response Relationship, Drug, Histamine pharmacology, Humans, Arthritis, Rheumatoid immunology, Cimetidine pharmacology, Diphenhydramine pharmacology, Lymphocyte Activation drug effects, Phytohemagglutinins pharmacology, Psoriasis immunology
- Abstract
Peripheral blood lymphocytes of patients with rheumatoid arthritis, psoriatic arthritis and healthy controls, isolated by a ficoll density gradient, were incubated with different concentrations of histamine, cimetidine and diphenhydramine (DPH) and simultaneously stimulated with phytohemagglutinine (PHA). In comparison to healthy control persons the lymphoproliferation rate in arthritic patients could be increased about 20% by addition of the histamine-blocking agents cimetidine and DPH. This suggests the existence of in vivo histamine-induced suppressor cells in arthritic patients, which can be inhibited by receptor blocking agents.
- Published
- 1984
19. The present status of the control of airborne infections.
- Author
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STIX R
- Subjects
- Humans, Infection Control, Infections
- Published
- 1947
20. A PUBLIC HEALTH PROGRAM FOR CHILDREN WITH HEART DISEASE OR RHEUMATIC FEVER.
- Author
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BROWNELL KD and STIX RK
- Subjects
- Child, Humans, New York, Heart Defects, Congenital, Heart Diseases, Public Health, Rheumatic Fever, Rheumatic Heart Disease, Statistics as Topic
- Published
- 1963
- Full Text
- View/download PDF
21. The place of fertility control in public health.
- Author
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STIX RK
- Subjects
- Humans, Contraception, Public Health
- Published
- 1946
22. Abortion and the Public Health.
- Author
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Stix RK and Wiehl DG
- Published
- 1938
- Full Text
- View/download PDF
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