Completed Collaboration & Service Projects

 


 

1) Relationship between structural Dynamics and Sequence Evolution; Application to molecular machined such as bacterial chaperonons and HSP70 chaperones

PI: I. Bahar, University of Pittsburgh  Collaborators: Amnon Horovitz, Weizmann Institute; R. Altman, Stanford University; L. Gierasch, University of Massachusetts.

aanmWe have expanded the scope of this project to include Dr. Amnon Horovitz from Weizmann Institute, Israel, an expert in the allostery of chaperonins as well as sequence co-evolution. We are exploring the sequence->structure->dynamics->function mapping of allosteric proteins. Our overarching goal is to establish the computational methodology for simulating the machinery of biomolecular systems on the order of Megadaltons. The molecular machines that we selected for developing and implementing the methodology are bacterial chaperonins (GroEL), molecular chaperones (E. coli Hsp70 DnaK) and their co-chaperones. We study the ATP-regulated mechanisms of intersubunit (GroEL) or interdomain (DnaK) communication, as well as the role of co-chaperonins or co-chaperones in modulating the conformational changes occurring along the allosteric cycle of these machines.

In Year 3 we made progress in quantitative analysis of the evolution of proteins, and in particular in assessing the relationship, if any, between their structural dynamics and sequence evolution. In a recent study performed toward this goal (Mao et al., 2015), we carried out a systematic comparative analysis of current methods for analyzing sequence co-evolution and we proposed a hybrid model that yields high performance. The two metrics for performance were the elimination of false positives between non-interacting proteins, and the verification of tertiary contacts between co-evolving pairs. The methods used in this newly published (that acknowledged MMBioS support) are currently being implemented into our ProDy API.

Publications Resulting from This Work

    • General IJ, Liu Y, Blackburn M, Mao W, Gierasch LM, Bahar I (2014) ATPase subdomain IA is a mediator of interdomain allostery in Hsp70 molecular chaperones PLoS Comp Bio 10: e1003624 PMID: 24831085, PMC4022485
    • Mao W, Kaya C, Dutta A, Horovitz A, Bahar I (2015) Comparative Study of the Effectiveness and Limitations of Current Methods for Detecting Sequence Coevolution Bioinformatics 31: 1929-37 PMID: 25697822; PMC4481699

Completed


 

2) Membrane Proteins Structure and Dynamics Consortium (MPSDC) Computational Core, and AMPA receptors structural dynamics

PI: I. Bahar, University of Pittsburgh   Collaborators: B. Roux and E. Perozo, University of Chicago; K. Schulten and E. Tajkhorshid, University of Illinois at Urbana-Champaign;  I. Greger, University of Cambridge, UK; H. Weinstein, Cornell University

To assess whether ANM-predicted AMPAR NTD pivoting motions can occur in cellulo, the Greger lab mutated K262 to cysteine (K262C), hypothesizing that this mutation would trap the NTD tetramer in a closed conformer via formation of a disulfide bridge. GluA2 wild type (WT) and K262C mutant were expressed in HEK293 cells and protein extracts were analyzed by Western blot. The results showed that our hypothesis was valid, thus lending support to computational predictions. We further examined the dynamics of AMPAR and NMDAR to observe that the differ along a few slow modes only (Figure 1). Overall, our data provided a first glimpse into the dynamic spectrum of AMPAR and NMDARs and delineated conserved mechanisms underlying allosteric communication in iGluRs.

csp2Figure 1. AMPAR to NMDAR conformations differ by rearrangements along a few soft modes. The dark blue bars show the overlap (correlation cosine) between ANM modes of AMPAR and the structural difference vector between AMPAR and NMDAR; the red curve displays the cumulative overlap; the dotted green curve shows the expected cumulative overlap if the modes were equally contributing each. The inset shows the initial structures of AMPAR (left) and NMDAR (right).

Publications Resulting from This Work

    • Krieger J, Bahar I, Greger IH (2015) Structure, Dynamics, and Allosteric Potential of Ionotropic Glutamate Receptor N-Terminal Domains Biophys J109: 1136-48 PMID: 26255587, PMC4576161
    • Dutta A, Krieger J, Lee JY, Garcia-Nafria J, Greger IH, Bahar I (2015) Cooperative Dynamics of Intact AMPA and NMDA Glutamate Receptors: Similarities and Subfamily-Specific Differences Structure23: 1692-1704 PMID: 26256538, PMC4558295
    • Dutta A, Shrivastava IH, Sukumaran M, Greger IH, Bahar I (2012) Comparative dynamics of NMDA- and AMPA-Glutamate receptor N-terminal domains Structure20: 1838-49 PMID: 22959625, PMC3496038

Completed

 


 

3) Integration, prediction, and generation of mixed mode information using graphical models, with applications to protein-protein interactions

PIs: C. J. Langmead, Carnegie Mellon University Collaborators: C. Bailey-Kellog, Dartmouth University; N. Ramakrishnan, Virginia Tech;  A. Friedman, Purdue University

This C&SP was an NSF-funded project titled: “Integration, prediction, and generation of mixed mode information using graphical models, with applications to protein-protein interactions” (NSF IIS-0905193). It was collaboration between Carnegie Mellon University, Dartmouth College, Virginia Tech, and Purdue. The project concluded in August, 2013. The focus of this C&SP was to develop novel probabilistic graphical models for modeling protein-protein interactions. Three broad classes of techniques were developed to integrate attribute-value and relational information, integrate statistical and physical information, and utilize probabilistic models generatively. The research has resulted in new graphical models, new algorithms, and the application of those algorithms to the modeling of protein structures and dynamics, including protein-protein interactions. Specific accomplishments include: the first optimal algorithm for learning regularized undirected graphical models of protein sequences; the first method for performing binding free energy calculations for protein-protein interactions via graphical models; the first probabilistic graphical models of molecular dynamics; the first algorithm for learning the parameters of molecular mechanics force fields by minimizing functionals over Boltzmann distributions; the first undirected model of distributions on the hypersphere (for modeling distributions over angles); a game-theoretic method for modeling the emergence of drug resistance-causing mutations in proteins; new algorithms for learning semi- and non-parametric distributions; and a new class of regression models for predicting binding free energies. The project resulted in seventeen publications, three Ph.D. dissertations, and one patent. Primary applications of this research include computer-aided drug design and computer-aided protein engineering. Secondary applications of this work include techniques for identifying functionally important residues mediating binding and allostery.

Publications Resulting from This Work

    • Kamisetty H, Ghosh B, Langmead CJ, Bailey-Kellogg C (2014) Learning Sequence Determinants of Protein: protein Interaction Specificity with Sparse Graphical Models Res Comput Mol Biol8394:129-143 PMID: 25414914, PMC4235964.
    • Kamisetty H, Ghosh B, Langmead CJ, Bailey-Kellogg C (2015) Learning sequence determinants of protein: protein interaction specificity with sparse graphical models J Comput Biol22:474- 86 PMID: 25973864, PMC4449715

Completed


 

4) Calcium entry and transmitter release at the neuromuscular junction

PIs: T. Bartol, T. Sejnowski, Salk Institute; R. Laghaei, G. Hood, M. Dittrich, Pittsburgh Supercomputing Center  Collaborator: S. Meriney, University of Pittsburgh

Communication between cells in the nervous system (synaptic transmission) underlies all complex behaviors, is often disrupted in neurological disease, and is a focus for therapeutic intervention. Synapses work by releasing chemical transmitter from a region called the active zone. In this project we synergistically combine computer simulation using MCell (Dittrich lab) and synaptic anatomy, physiology, and Ca2+ imaging (Meriney lab) to investigate the structure and function of synapses. We have previously completed and published a baseline model of frog neuromuscular junction (NMJ) active zone structure and function (Dittrich et al., 2013). Based on this model we have recently investigated several possible mechanisms underlying short-term synaptic facilitation at the frog NMJ (Ma et al., 2015). Our study showed that a vesicle release mechanism in which a second set of Ca2+ sensor sites was responsible for facilitation provided good overall agreement with our experimental constraints. In addition, we used our MCell modeling approach combined with Ca2+ imaging, pharmacological Ca2+ channel block, and postsynaptic recording to show that release of individual synaptic vesicles at the frog NMJ is predominately triggered by Ca2+ ions entering the nerve terminal through the nearest open calcium channel. This work has recently been published (Luo et al., 2015). Building on these insights, we are currently expanding our investigation to the role of detailed active zone structure in synapse function with a focus on neuromuscular diseases such as Lambert-Eaton Myasthenic Syndrome.

Publications Resulting from This Work

    • Tarr TB, Dittrich M, Meriney SD (2013) Are unreliable release mechanisms conserved from NMJ to CNS? Trends Neurosci36(1):14-22. PMID: 23102681, PMC4076818
    • Meriney SD, Dittrich M (2013) Organization and function of transmitter release sites at the neuromuscular junction J Physiol591(13):3159-65. PMID: 23613535, PMC3717219
    • Dittrich M, Pattillo JM, King JD, Cho S, Stiles JR, Meriney SD (2013) An excess-calcium-binding-site model predicts neurotransmitter release at the neuromuscular junction Biophys J104(12):2751-63 PMID: 23790384, PMC3686347
    • Ma J, Kelly L, Ingram J, Price TJ, Meriney SD, Dittrich M (2015) New insights into short-term synaptic facilitation at the frog neuromuscular junction J Neurophysiol113:71-87 PMID: 25210157, PMC4294575
    • Luo F, Dittrich M, Cho S, Stiles JR, Meriney SD (2015) Transmitter release is evoked with low probability predominately by calcium flux through single channel openings at the frog neuromuscular junction J Neurophysiol113:2480-9 PMID: 25652927, PMC4416571

Completed


 

6) Distance-dependent structure and function of neuronal dendrites

PIs: T. Sejnowski and T. Bartol, The Salk Institute Collaborator: K. Harris, University of Texas at Austin

The purpose of this CS&P is to collaborate with Kristen Harris' lab to create realistic MCell models of dendritic structure and function.

MCell can simulate the diffusion and interaction of molecules involved in biochemical signaling pathways within the 3D subcellular structure of cells. To do so, surface meshes used to represent cell membranes and subcellular structures must meet very strict geometric standards (e.g. water-tight, non-intersecting, manifold). We help create realistic MCell models of CA1 dendritic spines, in particular to learn how more the accurate serial section tomographic reconstructions of core organelles impact simulations of molecular signaling within dendritic spines.

Recently two papers were published relating to this C&SP. The first paper, published in Frontiers in Synaptic Neuroscience, reported on reconstitution of calcium dynamics in dendritic spines. Nine parameters of the model were optimized within realistic experimental limits by a process that compared results of simulations to published data. Simulations in the optimized model reproduce the timing and amplitude of Ca(2+) transients measured experimentally in intact neurons. Thus, the characteristics of individual isolated proteins determined in vitro could accurately reproduce the dynamics of experimentally measured Ca(2+) transients in spines. The second paper, published in eLife, found that dendritic spines that receive input from the same axon are the same size. This finding allowed estimation of the variability of synaptic plasticity and we found that the amount of information stored at synapses is approximately 4.7 bits, which is an order of magnitude larger than previous estimates.

Publications Resulting from This Work

    • Edwards J, Daniel E, Kinney J, Bartol T, Sejnowski T, Johnston D, Harris K, Bajaj C (2014) VolRoverN: enhancing surface and volumetric reconstruction for realistic dynamical simulation of cellular and subcellular functionNeuroinformatics12:277-89 PMID: 24100964, PMC4033674
    • Kinney JP, Spacek J, Bartol TM, Bajaj CL, Harris KM, Sejnowski TJ (2013) Extracellular sheets and tunnels modulate glutamate diffusion in hippocampal neuopil J Comp Neurol521: 448-64 PMID: 22740128, PMC3540825
    • Bartol TM, Keller DX, Kinney JP, Bajaj CL, Harris KM, Sejnowski TJ, Kennedy MB (2015) Computational reconstitution of spine calcium transients from individual proteins Front Synaptic Neurosci7: 17 PMID: 26500546, PMC4595661
    • Bartol TM, Bromer C, Kinney J, Chirillo MA, Bourne JN, Harris KM, Sejnowski TJ (2015) Nanoconnectomic upper bound on the variability of synaptic plasticity Elife4.pii: e10778 PMID: 26618907, PMC4737657

Completed


 

9) Large-scale electron microscopy of calcium-imaged neuron populations

PIs: A. Wetzel and G. Hood, Pittsburgh Supercomputing Center Collaborator: D. Bock, Janelia Farm Research Campus, Howard Hughes Medical Institute

During the last year of this project, the biological focus of Bock's work has shifted from localized mouse brain circuits to the mapping of complete drosophila brains with an emphasis on visual circuits. This change did not affect the relation of our collaboration which is focused on the technical aspects of maximum scale serial-TEM data assembly.

Bock's group has improved the throughput of their automated 4 camera TEM system. Although the cameras and their raw data rates are unchanged new mechanisms have been tested in the last year for automatic sample cassette loading. This has substantially improved the ratio of imaging to sample exchange time. The long term goal for this new sample loading method is to enable continuous runs up to 2 weeks for datasets of 15,000 sections and >50 TBytes. Individual sections are imaged in arrays of ~20,000 5 MPixel tiles, typically 112 columns by 196 rows, with a unique stepping pattern to accommodate constraints on the spacing of the optical cameras and lenses. One difficult aspect of these data is the use of the small 5 MPixel tiles which, particularly in the case of hierarchical registration methods, become limited by poor performance of large filesystems when processing small files.

Regular production of datasets at this scale would be difficult to register using our previous method of raw data transmission to the PSC followed by the return of aligned results to Janelia Farm. Therefore we are preparing for Bock's team to run our alignment codes on the large 6,144 core Janelia Farm computing cluster. Wetzel and Hood were part of an April 1-11, 2015 Janelia Farm image registration Hackathon organized by Stephan Saafeld's group. During that trip we were able to test both the AlignTK and SWiFT codes on the Janelia systems using data from Bock's work as well as many others from Janelia and elsewhere (i.e. Winfried Denk and our C&SP10 collaborator Jeff Lichtman). We were also able to adapt a key component of the SWiFT method for use within one of the Janelia registration codes and found that it greatly improved the reliability of registration on difficult regions of Bock's data.

Publications Resulting from This Work

    • Bock DD, Lee WC, Kerlin AM, Andermann ML, Hood G, Wetzel AW, Yurgenson S, Soucy ER, Kim HS, Reid RC (2011) Network anatomy in vivo physiology of visual cortical neurons Nature471: 177-82 PMID: 21390124, PMC3095821

Completed


 

10) Advancing high-throughput thin-section scanning EM to study relationships between neuronal circuit structure and function

PIs: A. Wetzel and G. Hood, Pittsburgh Supercomputing Center Collaborator: J. Lichtman, Harvard University

We have continued work on automated alignment of serial SEM image sets produced using tape collecting ultramicrotome sectioning and wafer mounted imaging procedures developed at Harvard. The primary datasets have been the 16,000 section zebrafish stack, which is the focus of a new CS&P project with the Engert lab and the 114 TB lateral geniculate nucleus (LGN) volume that was previously acquired and is the focus of this C&SP.

The different requirements and resolutions of these datasets have highlighted the need for a variety of approaches to handle different types of biological content and different types of image characteristics. The large number of tape cracks in the LGN dataset has been particularly difficult since the cracks from any section to its neighbors interfere with one another during the alignment process. Even though the detailed cracking pattern is different on each section the fact that most of the cracks have similar sizes and orientations means that they tend to pull the alignment such that we have seen errors on the order of 1 per 1000 sections. We are currently working on a strategy to produce a low-resolution map of the cracks, 1/4th scale, that can then be used to constrain the SWiFT alignment process.

We are entering a new and much higher-throughput phase of EM data capture. Previous datasets, including the zebrafish and LGN, were captured over several months using a Zeiss Merlin microscope and pixel rates up to 20MHz. Lichtman's team recently installed the first Zeiss 61-beam 1.2 Gpixel/sec scanning microscope. This microscope is now being tested with large single sections in the 400GB/section range, which are captured in a hexagonal mosaic pattern. These data will require a substantial number of registration code adaptations. We will test speed and accuracy of these modifications once the new equipment is producing serial spans of at least 100 sections, which is where the power of the SWiFT approach is most useful.

Publications Resulting from This Work

    • Morgan JL, Berger DR, Wetzel AW, Lichtman JW (2016) The fuzzy logic of network connectivity in mouse visual thalamus Cell165: 192-206 PMID: 27015312, PMC4808248
    • Hildebrand DGC, Torres RM, Choi W, Tran Minh Quan, Arthur Willis Wetzel, George ScottPlummer, Ruben Portugues, Isaac Henry Bianco, Owen Randlett, Stephan Saalfeld, Alex Baden, Kunal Lillaney, Randal Burns,Joshua Tzvi Vogelstein, Won-Ki Jeong, Jeff William Lichtman, Florian Engert (2016) Whole-brain serial-section electron microscopy in larval zebrafish Nature, revised version in preparation.

Completed


 

11) Morphological and regulatory models of neuronal differentiation

PIs: H Busch and M Böerries, University of Freiburg Collaborators: R Murphy and G Rohde, Carnegie Mellon University

The goal of this project is to develop a spatiotemporal generative model of the changes in cell size, shape and subcellular organization that occur during the differentiation of PC12 cells induced by nerve growth factor. This model will be related to a regulatory model constructed from parallel measurements of RNA expression over the time course of differentiation. The results are expected to lead to the identification of gene expression changes that lead to specific morphological changes and to test their importance for the differentiation process.

Publications Resulting from This Work

    • Johnson GR, Bierschenk I, Nitschke R, Boerries M, Busch H, and Murphy RF. (2017) Image-derived Models of Cell Organization Changes During Differentiation of PC12 Cells, submitted.

Completed


12) Generative models of plant organelle distribution and differentiation

PI: K Palme, University of Freiburg Collaborator: R Murphy, Carnegie Mellon University

Plants exhibit the ability to undergo dramatic dedifferentiation and redifferentiation such that both protoplasts (cells extracted from mature plants) and microspores (cells that arise during gametogenesis) can give rise to mature plants under the right circumstances. The goal of this project is to identify the specific changes in protein expression and localization that are associated with such processes, in part by building spatiotemporal generative models directly from images and movies of cells obtained by light microscopy.

Publications Resulting from This Work

    • Johnson GR, Kangas JD, Dovzhekno A, Trojok R, Voigt K, Majarian TD, Palme K and Murphy RF. (2017) A Method for Characterizing Phenotypic Changes in Highly Variable Cell Populations and its Application to High Content Screening of Arabidopsis thaliana Protoplasts. Cytometry Part A, in press.

Completed


13) Microtubule pattern analysis and drug sensitivity

PI: P Giannakakou, Cornell University Collaborator: R Murphy, Carnegie Mellon University

The goal of this new project is to determine whether and how the distributions of microtubules differ between tumor cells sensitive and resistant to drugs that act on microtubules. Initial work has focused on training systems to recognize these patterns, with a complicating factor being the large differences in cell size, shape and unperturbed microtubule pattern between cells from different tumors. Preliminary results indicate that three broad but distinct patterns can be distinguished, corresponding to untreated cells, drug-treated resistant cells, and drug-treated sensitive cells. Future work will focus on using generative models to remove variation due to cell size and shape. Ultimately, this would be used to determine whether a particular tumor is likely to be resistant to a particular drug and thereby to choose an appropriate drug for that tumor.

Completed


 

14) Actin filament patterns and cell motility

PI: J Theriot, Stanford University Collaborators: G Rohde and D Slepčev, Carnegie Mellon University

This is a new project, the goal of which is to quantitatively describe the patterns of actin filaments and related proteins as they relate to cell motility in neutrophils. The project aims to use recently developed methods for measuring the similarity between such patterns, as well as transport-based method for finding correlation between intensity patterns, to decode relationships between different proteins as cells move. Results are expected is to help characterize cell motion as a function of the subcellular protein patterns.

Completed


 

21) Functional significance of the dynamics of AMPAR extracellular region

Collaborating Investigators: Ingo H. Greger, MRC Lab of Molecular Biology, Ivet Bahar, QingDe Wang, University of Pittsburgh, Tom M. Bartol, Terry J. Sejnowski, Salk Institute

Ionotropic glutamate receptor (iGluRs) are ligand-gated ion channels that allow for the flow of cations into the postsynaptic cell in response to glutamate binding, thus regulating neurotransmission upon depolarization of the cell membrane. Among iGluR subfamilies, AMPAR and NMDAR play a key role in learning and memory, and in particular the AMPAR is essential to rapid neurotransmission and synaptic plasticity. The Greger and Bahar labs have been productively collaborating in recent years on AMPAR dynamics, first using the NTD dimer structures and more recently the intact tetrameric structures. These studies demonstrated that the NTD domains exhibit structural flexibilities comparable to those of AMPAR NTDs. Furthermore, the global modes of motions predicted by ANM (or ProDy) revealed the propensity of homotetrameric AMPAR to assume more compact forms similar to NMDARs. The validity of these modes of motions were confirmed by cross-linking experiments between NTD sites predicted by ANM to come into close proximity. In the new term, we will first adopt ANM-based analysis to characterize the mode spectrum of the heterotetrameric AMPAR. ProDy analysis already revealed that the O ↔ N transition is enabled by a global ANM mode. We will characterize thoroughly the whole spectrum of motions and generate the energy landscape of Glu2/3 heterotetramer, using the recently introduced extension of coMD. Then we will focus on the ECR motions that induce a pore opening (or cooperative twisting) at the TMD and analyze the conformational events that enable the allosteric coupling between the ECR and the TMD with the help of accelerated MD simulations. In the next phase, we plan to examine the significance of GluA2/3 ECR flexibility in adapting to its interactions with auxiliary proteins such as cornichon homologs, TARPs or in forming clusters, which will be further tested/validated with structural and single-particle tracking methods in the Greger lab.

Publications Resulting from This Work

    • Krieger J, Bahar IGreger IH (2015) Structure, Dynamics, and Allosteric Potential of Ionotropic Glutamate Receptor N-Terminal Domains Biophys J 109: 1136-48 PMID: 26255587, PMC45761612
    • Dutta A, Krieger J, Lee JY, Garcia-Nafria J, Greger IHBahar I (2015) Cooperative Dynamics of Intact AMPA and NMDA Glutamate Receptors: Similarities and Subfamily-Specific Differences Structure 23: 1692-1704 PMID: 26256538, PMC4558295
    • Dutta A, Shrivastava IH, Sukumaran M, Greger IHBahar I (2012) Comparative dynamics of NMDA- and AMPA-Glutamate receptor N-terminal domains Structure 20: 1838-49 PMID: 22959625, PMC3496038
    • Lee JY, Krieger J, Herguedas B, García-Nafría J, Dutta A, Shaikh SA, Greger IH, Bahar I. (2019) Druggability Simulations and X-ray Crystallography Reveal a Ligand-binding Site in the GluA3 AMPA Receptor N-terminal Domain. Structure 27: 241-252. 
    • Krieger J, Lee JY, Greger IH, Bahar I. (2018) Activation and Desensitization of Ionotropic Glutamate Receptors by Selectively Triggering Pre-existing Motions. Neurosci Lett 700: 22-29 PMID: 29481851 PMCID: 6107436

Completed


 

25) Efficient parallel sampling at multiple scales using the weighted ensemble strategy

Collaborating Investigators: Lillian T. Chong, University of Pittsburgh, Daniel Zuckerman, Oregon Health and Science University

There is a "silicon ceiling" that ultimately limits many, if not most, types of dynamical biological simulations. That is, even the world's most powerful computers cannot generate sufficiently long simulations, whether for atomistic models of proteins or for realistic models of cell behavior. In many cases, the key events may occur beyond simulation timescales - such as protein folding, conformational transitions of proteins, assembly of protein complexes, or transitions of cell behavior from healthy to pathological states. The work in this C&SP will continue the development of WESTPA, a tool for controlling other software tools: it orchestrates up to thousands of trajectories run natively by other software at any scale (e.g., Gromacs, Amber, BioNetGen, MCell) using a "weighted ensemble" strategy. Not only does WESTPA parallelize the use of dynamics engines - but because of the statistical process by which trajectories are added and removed, WESTPA can obtain estimates of key kinetic as well as equilibrium observables in significantly less computing time than would be required in ordinary parallelization. Our aims are to improve the ease of use and interoperability of WESTPA; to improve its performance and reliability; to demonstrate the effectiveness of WESTPA through a series of "showcase" examples from molecular to cellular scale using a variety of dynamics engines; and to improve instructional materials based on the showcase examples.

Publications Resulting from This Work

    • Zwier MC, Adelman JL, Kaus JW, Pratt AJ, Wong KF, Rego NB, Suárez E, Lettieri S, Wang DW, Grabe M, Zuckerman DM, Chong LT (2015) WESTPA: an interoperable, highly scalable software package for weighted ensemble simulation and analysis J Chem Theory Comput 11: 800-809. PMID: 26392815; PMC4573570
    • Suárez E, Pratt AJ, Chong LT, Zuckerman DM (2016) Estimating First Passage Time Distributions from Weighted Ensemble Simulations and non-Markovian Analyses Protein Science 25: 67-78. PMID: 26131764; PMC4815309
    • Suárez E, Pratt AJ, Chong LT, Zuckerman DM (2016) Estimating First Passage Time Distributions from Weighted Ensemble Simulations and non-Markovian Analyses Protein Science 25: 67-78. PMID: 26131764; PMC4815309

Completed


 

28) Dynamic modulation of interferon binding affinity as a mechanism to regulate interferon receptor signaling

Collaborating Investigators: Gideon Schreiber, Weizmann Institute, Ivet Bahar, James R. Faeder, University of Pittsburgh

Type I interferons (IFNs) are multifunctional cytokines that mediate/induce diverse cellular responses, including both innate and adaptive immune responses, stimulation of antiviral responses, and cancer surveillance, upon forming a ternary complex with two surface receptors, IFNAR1 and IFNAR2. The activities of IFN-a subtypes correlate with their affinities to bind to IFNAR1 and IFNAR2. While the Schreiber lab made seminal contributions to understanding the molecular basis of IFNARs, the mechanism of regulation of differential IFN activities through interactions with IFNAR1 and 2, remains unclear. Our integrated computational (TR&D1) and experimental preliminary studies point to the significance of the intrinsic dynamics in modulating binding affinity. We adopted a closely integrated computational/experimental strategy that yielded promising results, which we are currently further pursuing and that illustrate the adaptability of proteins to different bound states or to sequence variations/mutations via their softest modes of motion. Further cross-linking, fluorescence quenching and gene induction experiments will be conducted in the Schreiber lab, in close coordination with TR&D1 computational studies at the Bahar lab.

Publications Resulting from This Work

  • Li H, Sharma N, General IJ, Schreiber G, Bahar I. (2017) Dynamic Modulation of Binding Affinity as a Mechanism for Regulating Interferon Signaling. J Mol Biol 429: 2571-2589 PMID: 28648616 PMCID: 5545807

Completed

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