Computational Pharmaceutical Chemistry and Molecular Informatics Group

Permanent URI for this collection

https://researchdata.hhu.de/handle/entry/76

Our research focusses on understanding, predicting, and modulating biomolecular interactions from an atomistic perspective.

Browse

Now showing 1 - 20 of 23

Supporting Information for "Molecular mechanisms underlying single nucleotide polymorphism-induced reactivity decrease in CYP2D6"
(N/A, 2024-02) Becker, Daniel; Gohlke, Holger
Cytochrome P450 2D6 (CYP2D6) is one of the most important enzymes involved in drug metabolism. Genetic polymorphism can influence drug metabolism by CYP2D6 such that a therapy is seriously affected by under- or overdosing of drugs. However, a general explanation at the atomistic level for poor activity is missing so far. Here we show for the 20 most common single nucleotide polymorphisms (SNPs) of CYP2D6 that poor metabolism is driven by four mechanisms. We found in extensive all-atom molecular dynamics simulations that the rigidity of the I-helix (central helix), distance between central phenylalanines (stabilizing bound substrate), availability of basic residues on the surface of CYP2D6 (binding of Cytochrome P450 reductase), and position of arginine 132 (electron transfer to heme) are essential for an extensive function of the enzyme. These results were applied to SNPs with unknown effects and potential SNPs that may lead to poor drug metabolism were identified. The revealed molecular mechanisms might be important for other drug-metabolizing Cytochrome P450 enzymes.
MD simulation data for: "Molecular Mechanisms Underlying Medium-Chain Free Fatty Acid-regulated Activity of the Phospholipase PlaF from Pseudomonas aeruginosa"
(N/A, 2023-11) Gentile, Rocco; Schott-Verdugo, Stephan; Gohlke, Holger
PlaF is a membrane-bound phospholipase A1 from P. aeruginosa that is involved in remodeling membrane glycerophospholipids (GPLs) and modulation of virulence-associated signaling and metabolic pathways. Previously, we identified the role of medium-chain free fatty acids (FFA) in inhibiting PlaF activity and promoting homodimerization, yet the underlying molecular mechanism remained elusive. Here, we used unbiased and biased molecular dynamics simulations and free energy computations to assess how PlaF interacts with FFAs localized in the water milieu surrounding the bilayer or within the bilayer, and how these interactions regulate PlaF activity. Medium-chain FFAs localized in the upper bilayer leaflet can stabilize inactive dimeric PlaF, likely through interactions with charged surface residues as experimentally validated. Potential of mean force (PMF) computations indicate that membrane-bound FFAs may facilitate the activation of monomeric PlaF by lowering the activation barrier of changing into a tilted, active configuration. We estimated that the coupled equilibria of PlaF monomerization-dimerization and tilting at the physiological concentration of PlaF lead to the majority of PlaF forming inactive dimers when in a cell membrane loaded with decanoic acid (C10). This is in agreement with a suggested in vivo product feedback loop and GC-MS profiling results indicating that PlaF catalyzes the release of C10 from P. aeruginosa membranes. Additionally, we found that C10 in the water milieu can access the catalytic site of active monomeric PlaF, contributing to the competitive component of C10-mediated PlaF inhibition. Our study provides mechanistic insights into how medium-chain FFA may regulate the activity of PlaF, a potential bacterial drug target.
MD simulation data for: "The cyclophilin A binding loop of the capsid regulates the human TRIM5α sensitivity of nonpandemic HIV-1"
(N/A, 2023-11) Becker, Daniel; Münk, Carsten; Gohlke, Holger
All MD input structures, MD infiles, umbrella sampling files, and scripts that were used to analyze the umbrella sampling results are provided in this supporting repository.
Regulation of STING activity in DNA sensing by ISG15 modification
(N/A, 2023-09) Gertzen, Christoph; Kaiser, Jesko; Münk, Carsten; Gohlke, Holger
Molecular modeling and simulations suggest that ISGylation of K289 of STING is an important regulator of STING oligomerization
Supporting Information for "Enzyme adaptation to habitat thermal legacy shapes the thermal plasticity of marine microbiomes"
(N/A, 2023-01) Nutschel, Christina; Pfleger, Christopher; Dittrich, Jonas; Gohlke, Holger
The dataset contains: I. 3D structural models of esterases generated by a AlphaFold2-based workflow of ColabFold, II. predicted catalytic triads of esterases and their substrate accessibilities analyzed by CAVER 3.0.3 PyMOL Plugin, III. input and output files for molecular dynamics (MD) simulations performed by Amber21, and, IV. Tp-values of esterases predicted by Constraint Network Analysis (CNA). We added template scripts and a readme file for further explanations.
Supporting Information for "Loading and Co-Solvent-Triggered Release of Okanin, a C4 Plant Key Enzyme Inhibitor, into/from Functional Microgels"
(N/A, 2023-01) Dittrich, Jonas; Gohlke, Holger
The constantly growing world population leads to increasing demands for food, which challenges modern agriculture manifold. Pests, such as weeds, require the application of agrochemicals to increase crop yield. Due to the environmental impact of these potentially hazardous chemicals, the demand for more efficient formulations is increasing. Promising formulations consist of easily adaptable carriers from which controllable stimuli release the agrochemicals. Here, we investigated poly(N vinylcaprolactam) (pVCL)-based microgels as a potential carrier for okanin, an inhibitor of the C4 plant key enzyme phosphoenolpyruvate carboxylase, by combining experiments, molecular simulations, and free energy computations. Dynamic light scattering, scanning transmission electron and atomic force microscopy revealed that pVCL microgels collapse and rigidify upon the loading of okanin. The simulations identified loosely adsorbed okanin and tightly bound okanin mediating inter-chain crosslinks. With increasing okanin concentration, stacking interactions of okanin occur with adsorbed and bound okanin. These findings can explain the experimentally observed collapse and the rigidification of the microgels. Based on the atomistic insights, two poly(N vinylcaprolactam co glycidyl methacrylate) microgels were synthesized, for which a doubled loading capacity of okanin was found. Finally, we investigated the triggered release of okanin using the addition of green solvents as a stimulus. This work establishes a basis for the further optimization of pVCL-based microgels as a carrier for the delivery of polyphenolic agrochemicals.
TopEnzyme: A framework and database for structural coverage of the functional enzyme space
(N/A, 2022-12) Karel, van der Weg; Holger, Gohlke
TopEnzyme is a database of structural enzyme models created with TopModel and is linked to the SWISS-MODEL repository and AlphaFold Protein Structure Database to provide an overview of structural coverage of the functional enzyme space for over 200,000 enzyme models. It allows the user to quickly obtain representative structural models for 60% of all known enzyme functions. We assessed the models with TopScore and contributed 9039 good-quality and 1297 high-quality structures. Furthermore, we compared these models to AlphaFold2 models with TopScore and found that the TopScore differs only by 0.04 on average in favor of AlphaFold2. We tested TopModel and AlphaFold2 for targets not seen in the respective training databases and found that both methods create qualitatively similar structures. When no experimental structures are available, this database will facilitate quick access to structural models across the currently most extensive structural coverage of the functional enzyme space within Swiss-Prot.
Movie for: The architecture of the 10-23 DNAzyme and its implications for DNA-mediated catalysis
(N/A, 2022-09-19) Christoph, Gertzen; Jan, Borggräfe; Aldino, Viegas; Manuel, Etzkorn; Holger, Gohlke
Understanding the molecular features of catalytically active DNA sequences, so-called DNAzymes, is not only essential for our understanding of the fundamental properties of catalytic nucleic acids in general but may well be the key to unraveling their full potential via tailored modifications. Our recent findings contributed to the endeavor to assemble a mechanistic picture of DNA-mediated catalysis by providing high-resolution structural insights into the 10-23 DNAzyme (Dz) and exposing a complex interplay between the Dz´s unique molecular architecture, conformational plasticity, and dynamic modulation by metal ions as central elements of the DNA catalyst. To illustrate the sampled conformational space, the movie depicts one MD trajectory of the Dz:RNA complex.
Millisecond-long sampling for a comprehensive energetic evaluation of aqueous ionic liquid effects on amino acid interactions
(N/A, 2022-09-01) El Harrar, Till; Gohlke, Holger
The interactions of amino acid side-chains confer diverse energetic contributions and physical properties to a protein's stability and function. Various computational tools estimate the effect of changing a given amino acid on the protein's stability based on parametrized (free) energy functions. When parametrized for the prediction of protein stability in water, such energy functions can lead to suboptimal results for other solvents, such as ionic liquids (IL), aqueous ionic liquids (aIL), or salt solutions. However, to our knowledge, no comprehensive data is available describing the energetic effect of aIL on intramolecular protein interactions. Here, we present the most comprehensive set of potential of mean force (PMF) profiles of pairwise protein-residue interactions to date, covering 50 relevant interactions in water, the two biotechnologically relevant aIL [BMIM/Cl] and [BMIM/TfO], and [Na/Cl]. These results are based on a cumulated simulation time of > 1 ms. aIL and salt ions can weaken, but also strengthen, specific residue interactions by more than 3 kcal mol 1, depending on the residue pair, residue-residue configuration, participating ions, and concentration, necessitating considering such interactions specifically. These changes originate from a complex interplay of competitive or cooperative noncovalent ion-residue interactions, changes in solvent structural dynamics, or unspecific charge screening effects and occur at the contact distance but also at larger, solvent-separated distances. This data provides explanations at the atomistic and energetic level for complex IL effects on protein stability and should help improve the prediction accuracy of computational tools that estimate protein stability based on (free) energy functions.
Uncoupling of voltage- and ligand-induced activation in HCN2 channels by glycine inserts v2
(N/A, 2022) Sezin, Yüksel; Bonus, Michele; Pfleger, Christopher; Gohlke, Holger; Benndorf, Klaus; Kusch, Jana
Hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels are tetramers that generate electrical rhythmicity in special brain neurons and cardiomyocytes. The channels are activated by membrane hyperpolarization. The binding of cAMP to the four available cyclic nucleotide-binding domains (CNBD) enhances channel activation. We analyzed in the present study the mechanism of how the effect of cAMP binding is transmitted to the pore domain (PD). Our strategy was to uncouple the C-linker (CL) from the channel core by inserting one to five glycine residues between the S6 gate and the A’-helix (constructs 1G to 5G). We quantified in full-length HCN2 channels the resulting functional effects of the inserted glycines by current activation as well as the structural dynamics and statics using molecular dynamics simulations and Constraint Network Analysis (CNA). We show functionally that already in 1G the cAMP effect on activation is lost and that with the exception of 3G and 5G the concentration-activation relationships are shifted to depolarized voltages with respect to HCN2. The strongest effect was found for 4G. Accordingly, the activation kinetics were accelerated by all constructs, again with the strongest effect in 4G. The simulations reveal that the average residue mobility of the CL and CNBD domains is increased in all constructs and that the junction between the S6 and A’-helix is turned into a flexible hinge, resulting in a destabilized gate in all constructs. Moreover, for 3G and 4G, there is a stronger downward displacement of the CL-CNBD than in HCN2 and the other constructs, resulting in an increased kink angle between S6 and A’-helix, which in turn loosens contacts between the S4-helix and the CL. This is suggested to promote a downward movement of the S4-helix, similar to the effect of hyperpolarization. In addition, exclusively in 4G, the selectivity filter in the upper pore region and parts of the S4-helix are destabilized. The results provide new insights into the intricate activation of HCN2 channels.
Structural and mechanistic insights into bacterial phospholipase A involved in membrane phospholipid degradation and virulence
(N/A, 2022) Stephan, Schott-Verdugo; Holger, Gohlke; Renu, Batra-Safferling; Karl-Erich, Jaeger; Filip, Kovacic
Cells steadily adapt their membrane glycerophospholipid (GPL) composition to changing environmental and developmental conditions. While the regulation of membrane homeostasis via GPL synthesis in bacteria has been studied in detail, the mechanisms underlying the controlled degradation of endogenous GPLs remain unknown. Thus far, the function of intracellular phospholipases A (PLAs) in GPL remodeling (Lands cycle) in bacteria is not clearly established. Here, we identified the first cytoplasmic membrane-bound phospholipase A1 (PlaF) from Pseudomonas aeruginosa, which might be involved in the Lands cycle. PlaF is an important virulence factor, as the P. aeruginosa ΔplaF mutant showed strongly attenuated virulence in Galleria mellonella and macrophages. We present a 2.0-Å-resolution crystal structure of PlaF, the first structure that reveals homodimerization of a single-pass transmembrane (TM) full-length protein. PlaF dimerization, mediated solely through the intermolecular interactions of TM and juxtamembrane regions, inhibits its activity. The dimerization site and the catalytic sites are linked by an intricate ligand-mediated interaction network, which might explain the product (fatty acid) feedback inhibition observed with the purified PlaF protein. We used molecular dynamics simulations and configurational free energy computations to suggest a model of PlaF activation through a coupled monomerization and tilting of the monomer in the membrane, which constrains the active site cavity into contact with the GPL substrates. Thus, these data indicate the importance of the PlaF mediated GPL remodeling pathway for virulence and could pave the way for the development of novel therapeutics targeting PlaF.
Uncoupling of voltage- and ligand-induced activation in HCN2 channels by glycine inserts
(N/A, 2022) Sezin, Yüksel; Michele, Bonus; Christopher, Pfleger; Holger, Gohlke; Klaus, Benndorf; Jana, Kusch
Hyperpolarization-activated cyclic nucleotide-modulated (HCN) channels are tetramers that generate electrical rhythmicity in special brain neurons and cardiomyocytes. The channels are activated by membrane hyperpolarization. The binding of cAMP to the four available cyclic nucleotide-binding domains (CNBD) enhances channel activation. We analyzed in the present study the mechanism of how the effect of cAMP binding is transmitted to the pore domain (PD). Our strategy was to uncouple the C-linker (CL) from the channel core by inserting one to five glycine residues between the S6 gate and the A’-helix (constructs 1G to 5G). We quantified in full-length HCN2 channels the resulting functional effects of the inserted glycines by current activation as well as the structural dynamics and statics using molecular dynamics simulations and Constraint Network Analysis (CNA). We show functionally that already in 1G the cAMP effect on activation is lost and that with the exception of 3G and 5G the concentration-activation relationships are shifted to depolarized voltages with respect to HCN2. The strongest effect was found for 4G. Accordingly, the activation kinetics were accelerated by all constructs, again with the strongest effect in 4G. The simulations reveal that the average residue mobility of the CL and CNBD domains is increased in all constructs and that the junction between the S6 and A’-helix is turned into a flexible hinge, resulting in a destabilized gate in all constructs. Moreover, for 3G and 4G, there is a stronger downward displacement of the CL-CNBD than in HCN2 and the other constructs, resulting in an increased kink angle between S6 and A’-helix, which in turn loosens contacts between the S4-helix and the CL. This is suggested to promote a downward movement of the S4-helix, similar to the effect of hyperpolarization. In addition, exclusively in 4G, the selectivity filter in the upper pore region and parts of the S4-helix are destabilized. The results provide new insights into the intricate activation of HCN2 channels.
Functional and structural characterization of interactions between opposite subunits in HCN pacemaker channels
(N/A, 2021-08-05) Kondapuram, Mahesh; Frieg, Benedikt; Yüksel, Sezin; Schwabe, Tina; Sattler, Christian; Lelle, Marco; Schweinitz, Andrea; Schmauder, Ralf; Benndorf, Klaus; Gohlke, Holger; Kusch, Jana
Hyperpolarization-activated and cyclic nucleotide (HCN) modulated channels are tetrameric cation channels. In each of the four subunits, the intracellular cyclic nucleotide-binding domain (CNBD) is coupled to the transmembrane domain via a helical structure, the C-linker. High-resolution channel structures suggest that the C-linker enables functionally relevant interactions with the opposite subunit, which might be critical for coupling the conformational changes in the CNBD to the channel pore. We combined mutagenesis, patch-clamp technique, confocal patch-clamp fluorometry, and molecular dynamics simulations to show that residue K464 of the C-linker is essential for stabilizing the closed state of the mHCN2 channel by forming interactions with the opposite subunit. MD simulations revealed that both cAMP and K464E induce a rotation of the intracellular domain relative to the channel pore, weakening the autoinhibitory effect of the unoccupied CL-CNBD region. The adopted poses are in excellent agreement with structural results.
Simulation data for "Substrate access mechanism in a novel membrane-bound phospholipase A of Pseudomonas aeruginosa concordant with specificity and regioselectivity"
(N/A, 2021-07-12) Ahmad, Sabahuddin; Strunk, Christoph; Schott-Verdugo, Stephan; Jaeger, Karl-Erich; Kovacic, Filip; Gohlke, Holger
PlaF is a cytoplasmic membrane-bound phospholipase A1 from Pseudomonas aeruginosa that alters the membrane glycerophospholipid (GPL) composition and fosters the virulence of this human pathogen. PlaF activity is regulated by a dimer-to-monomer transition followed by tilting of the monomer in the membrane. However, how substrates reach the active site and how the characteristics of the active site tunnels determine the activity, specificity, and regioselectivity of PlaF for natural GPL substrates has remained elusive. Here, we combined unbiased and biased all-atom molecular dynamics (MD) simulations and configurational free energy computations to identify access pathways of GPL substrates to the catalytic center of PlaF. Our results map out a distinct tunnel through which substrates access the catalytic center. PlaF variants with bulky tryptophan residues in this tunnel revealed decreased catalysis rates due to tunnel blockage. The MD simulations suggest that GPLs preferably enter the active site with the sn-1 acyl chain first, which agrees with the experimentally demonstrated PLA1 activity of PlaF. We propose that the acyl chain-length specificity of PlaF is determined by the structural features of the access tunnel, which results in favorable free energy of binding of medium-chain GPLs. The suggested egress route conveys fatty acid products to the dimerization interface and, thus, contributes to understanding the product feedback regulation of PlaF by fatty acid-triggered dimerization. These findings open up opportunities for developing potential PlaF inhibitors, which may act as antibiotics against P. aeruginosa.
Supporting Information for "F/G-Region Rigidity is Inversely Correlated to Substrate Promiscuity of Human CYP Isoforms Involved in Metabolism"
(N/A, 2021-07-13) Becker, Daniel; Bharatam, Prasad; Gohlke, Holger
Of 57 human Cytochrome P450 (CYP) enzymes, twelve metabolize 90% of xenobiotics. To our knowledge, no study has addressed the relation between enzyme dynamics and substrate promiscuity for more than three CYPs. Here, we show by constraint dilution simulations with the Constraint Network Analysis for the twelve isoforms that structural rigidity of the F/G-region is significantly inversely correlated to the enzymes’ substrate promiscuity. This highlights the functional importance of structural dynamics of the substrate tunnel.
TopProperty dataset
(N/A, 2021-07-07) Mulnaes, Daniel; Schott-Verdugo, Stephan; Koenig, Filip; Gohlke, Holger
Transmembrane proteins (TMPs) are critical components of cellular life. However, due to experimental challenges, the number of experimentally resolved TMP structures is severely underrepresented in databases compared to their cellular abundance. Prediction of (per-residue) features such as transmembrane topology, membrane exposure, secondary structure, and solvent accessibility can be a useful starting point for experimental design or protein structure prediction, but often requires different computational tools for different features or types of proteins. We present TopProperty, a meta-predictor that predicts all of these features for TMPs or globular proteins. TopProperty predictions are robust, especially for proteins with few sequence homologs, and significantly better than the evaluated state-of-the-art primary predictors on all quality metrics. TopProperty eliminates the need for protein type- or feature-tailored tools, specifically for TMPs. TopProperty is freely available as web server and standalone at https://cpclab.uni-duesseldorf.de/topsuite/.
TopDomain dataset v2.0
(N/A, 2021-05-01) Mulnaes, Daniel; Golchin, Pegah; Koenig, Filip; Gohlke, Holger
This is the TopDomain dataset v2.0 as described in: "TopDomain: Exhaustive Protein Domain Boundary Meta-Prediction Combining Multi-Source Information and Deep Learning" by Daniel Mulnaes, Pegah Golchin, Filip Koenig, and Holger Gohlke. This dataset contains three folder: dataset : Contains the full dataset and the TopDomain and TopDomainSeq predictions for the dataset training_set : Contains the fasta files of the TopDomain training set test_set : Contains the fasta files of the TopDomain test set Each fasta file has a header with three fields, in the following format: >system_name|domain_type|boundary_list Where: system_name contains the PDB ID and chain ID of the target protein domain_type contains target type, either single-domain or multi-domain boundary_list contains a list of residues annotated as domain boundaries separated by spaces, this field is empty for single-domain proteins as they have no domain boundaries The sequence is the fasta-sequence of the protein each line contains at most 100 residues of the protein sequence No protein in the test set shares more than 20% sequence identity to any protein in the training set.
Supporting Information for "Molecular modeling and simulations of DNA and RNA: DNAzyme as a model system"
(N/A, 2021-03-17) Gertzen, Christoph; Gohlke, Holger
Nowadays, the structural dynamics of DNA and RNA is accessible on an atomistic level on a micro- to millisecond time scale via molecular dynamics simulations. However, as DNA or RNA are highly charged molecules, performing such simulations is challenging as to the representation of intramolecular electrostatic interactions and those to solvent molecules and ions. This is particularly true for DNAzymes, where DNA and RNA backbones can come as close as 2.4 Å with their charged phosphate groups during the catalytic cycle. Here, we present tools to simulate the structural dynamics of a DNAzyme, with a focus on detailed instructions for the Amber suite of programs. Furthermore, we will show how to analyze metal ion binding within the DNAzyme.
Structural models of SFV Bet protein as SI for "Foamy Viruses, Bet, and APOBEC3 Restriction"
(N/A, 2021-03-18) Vasudevan, Ananda; Becker, Daniel; Luedde, Tom; Gohlke, Holger; Muenk, Carsten
Domain boundaries within the sequence of the SFV-chimpanzee Bet protein sequence were predicted with TopDomain. The two domains were modeled using the ROBETTA webserver. For each domain, ROBETTA generated five models. The best model was chosen based on the lowest Cα atom RMSD between the generated models and the corresponding ab initio modeled domain of PFV Bet. This consensus approach was also validated by TopScore in that domains with the lowest RMSD also have the lowest TopScore compared to the other four models. For the N-terminal domain, the overall TopScore is 0.47 and for the C-terminal one 0.45, indicating a moderate quality of the models. In the N-terminal domain, the helical part of the structure has low local TopScores, indicating good quality, but the N-terminal loops high local TopScores, indicating low quality. In the C-terminal domain, the β-strands have low local TopScores, indicating good quality, but helices and loops from residues 358–403 have high local TopScores, indicating low quality.
TopDomain dataset
(N/A, 2021) Mulnaes, Daniel; Golchin, Pegah; Koenig, Filip; Gohlke, Holger
This is the TopDomain dataset as described in: "TopDomain: Exhaustive Protein Domain Boundary Meta-Prediction Combining Multi-Source Information and Deep Learning" by Daniel Mulnaes, Pegah Golchin, Filip Koenig, and Holger Gohlke. This dataset contains two folder: training_set : Contains the fasta files of the TopDomain training set; test_set: Contains the fasta files of the TopDomain test set. Each fasta file has a header with three fields, in the following format: ">system_name|domain_type|boundary_list". Where: system_name contains the PDB ID and chain ID of the target protein; domain_type contains target type, either single-domain or multi-domain; boundary_list contains a list of residues annotated as domain boundaries separated by spaces, this field is empty for single-domain proteins as they have no domain boundaries. The sequence is the fasta-sequence of the protein, each line contains at most 100 residues of the protein sequence. No protein in the test set shares more than 20% sequence identity to any protein in the training set.

Browse

Recent Submissions