Scientific Program

Conference Series Ltd invites all the participants across the globe to attend 14th International Conference on Structural Biology Berlin, Germany.

Day 1 :

Keynote Forum

Henry M. Sobell

University of Rochester USA

Keynote: The centers of premeltons signal the beginning and ends of genes

Time : 09:30-10:15

Conference Series Structural Biology 2018 International Conference Keynote Speaker Henry M. Sobell photo
Biography:

Henry M Sobell completed his studies at Brooklyn Technical High School (1948-1952), Columbia College (1952-1956), and the University of Virginia, School of Medicine
(1956-1960). Instead of practicing clinical medicine, he then went to the Massachusetts Institute of Technology (MIT) to join Professor Alexander Rich in the Department
of Biology (1960-1965), where, as a Helen Hay Whitney Postdoctoral Fellow, he learned the technique of single crystal x-ray analysis. He then joined the Chemistry
Department at the University of Rochester, having been subsequently jointly appointed to both the Chemistry and Molecular Biophysics Departments (the latter at
the University of Rochester School of Medicine and Dentistry), becoming a full tenured Professor in both departments (1965-1993). He is now retired and living in the
Adirondacks in New York, USA.

Abstract:

Premeltons are examples of emergent structures (i.e., structural solitons) that arise spontaneously in DNA due to the presence of nonlinear excitations in its structure.  They are of two kinds: B-B (or A-A) premeltons form at specific DNA-regions to nucleate site-specific DNA melting.  These are stationary and, being globally nontopological, undergo breather motions that allow drugs and dyes to intercalate into DNA.  B-A (or A-B) premeltons, on the other hand, are mobile, and being globally topological, act as phase-boundaries transforming B- into A- DNA during the structural phase-transition.  They are not expected to undergo breather-motions.  A key feature of both types of premeltons is the presence of an intermediate structural-form in their central regions (proposed as being a transition-state intermediate in DNA-melting and in the B- to A- transition), which differs from either A- or B- DNA. Called beta-DNA, this is both metastable and hyperflexible – and contains an alternating sugar-puckering pattern along the polymer-backbone combined with the partial-unstacking (in its lower energy-forms) of every other base-pair.  Beta-DNA is connected to either B- or to A- DNA on either side by boundaries possessing a gradation of nonlinear structural-change, these being called the kink and the antikink regions.  The presence of premeltons in DNA leads to a unifying theory to understand much of DNA physical-chemistry and molecular-biology.  In particular, premeltons are predicted to define the 5’ and 3’ ends of genes in naked-DNA and DNA in active chromatin, this having important implications for understanding physical aspects of the initiation, elongation and termination of RNA-synthesis during transcription.  For these and other reasons, the model will be of broader interest to the general audience working in these areas.  The model explains a wide variety of data, and carries within it a number of experimental predictions – all readily testable – as will be described in my talk.

Keynote Forum

Yuri L. Lyubchenko

University of Nebraska Medical Center,USA

Keynote: Self-Assembly of Amyloid Proteins

Time : 10:30-11:15

Conference Series Structural Biology 2018 International Conference Keynote Speaker Yuri L. Lyubchenko photo
Biography:

Yuri L. Lyubchenko is Professor of Pharmaceutical Sciences at the University of Nebraska Medical Center, Omaha, NE, USA. His research focuses on understanding fundamental mechanisms underlying health and disease, which are key to developing new and more effective diagnostics and medications. This primarily basic research allows him not only identify new drug targets for small molecule drugs, it also leads to development of the nanotools and methods to discover novel approaches for diagnostic, treatment and disease prevention and to more rapidly determine their efficacy at the molecular level.

 

 

Abstract:

 

 

 

Figure 1. Schematic for the surface mediated amyloid aggregation model.

 

 

Statement of the Problem The amyloid cascade hypothesis is currently considered as the main model for a vast number of neurodegenerative diseases including Alzheimer’s, Parkinson’s, and Huntington’s diseases. Numerous studies have shown that amyloidogenic proteins are capable of spontaneous assembly into aggregates, and eventually form fibrillar structures found in amyloid or amyloid‐like deposits. However, there is a serious complication with translating current knowledge on amyloid aggregation in vitro to understand the aggregation process in vivo. If the critical concentration for the spontaneous aggregation of Aβ peptide in vitro is in the micromolar range, physiological concentrations of Aβ are in the low nanomolar range making impossible amyloids to assemble.

Methodology & Theoretical Orientation: We discovered a novel on-surface aggregation pathway that allows for spontaneous assembly of amyloid beta peptides at the physiological concentration range. We combined experimental studies involving single-molecule time-lapse AFM imaging with all-atom molecular dynamic simulations to characterize the on-surface self-assembly process of amyloid proteins. Experimental data demonstrate that on-surface aggregation occurs in the physiological range of concentrations of the proteins. Our combined experimental and computer modeling approaches demonstrate that the on-surface aggregation is a dynamic process, so the assembled aggregate can dissociate from the surface to the bulk solution. As a result, the dissociated oligomers can play roles of seeds for aggregation in the bulk solution, or start a neurotoxic effect such as phosphorylation of the tau protein to initiate its misfolding and aggregation. Both processes can lead to neurodegeneration.  

Conclusion & Significance: we posit that on-surface aggregation is the mechanism by which neurotoxic amyloid aggregates are produced under physiological conditions (Figure 1). A change in membrane properties leading to an increase in affinity of amyloid proteins to the membrane surface facilitates the assembly of stable oligomers. The proposed model is a significant departure from the current model as it directs the development of treatments and preventions towards approaches that control the cell membranes composition to prevent the on-surface aggregation process. ………………………………………………….                                                                      

  • Determination of 3D structures | Structural Biology in Cell Signalling
Location: Spreewald

Chair

Joachim Krebs

MPI for Biophysical Chemistry, Germany

Biography:

Peter B Stathopulos applies structural biology approaches to reveal molecular mechanisms driving calcium signaling processes in health and diseased states including
heart disease, cancer and immunodefi ciency. He integrates nuclear magnetic resonance spectroscopy and x-ray crystallography with a host of biophysical, chemical
biology and live cell methodologies to understand the relationship between structure and function of critical calcium signaling proteins. Ultimately, this structure-function
data is used for the rational identifi cation of new drug binding targets with the potential to modulate these pathways to maintain health or treat disease.

Abstract:

Calcium (Ca2+) is a universal signaling entity in eukaryotic cells mediating diverse processes such as the immune response,
hypertrophy, apoptosis, platelet aggregation and memory, to name a few. Th ese processes require a sustained elevation
of cytosolic Ca2+ levels which is facilitated by store operated Ca2+ entry (SOCE). SOCE is the process whereby endoplasmic
reticulum (ER) luminal Ca2+ depletion signals the opening of ion channels on the plasma membrane (PM) which facilitate
the movement of Ca2+ down the concentration gradient from the extracellular space into the cytosol. Th e principal molecules
that mediate SOCE include the ER resident stromal interaction molecule-1 (STIM1) and PM ORAI1 protein subunits which
assemble into a channel pore. Upon ER luminal Ca2+ depletion, STIM1 undergoes a destabilization coupled oligomerization
which leads to translocation of this Ca2+ sensor to ER-PM junctions where it couples to ORAI1 subunits and opens these PM
Ca2+ channels. Since the identifi cation of STIM1 and ORAI1 as the principal molecules driving SOCE, considerable progress
has been made elucidating their high-resolution structural mechanisms of action. Author will present available structural data
on the STIM1 Ca2+ sensing mechanism and how this regulator may complex to ORAI1 subunits. Th e coupling mechanism
revealed using soluble human STIM1 and ORAI1 fragments are congruent with the hexameric assembly elucidated in the D.
melanogaster crystal structure. Finally, author will present unpublished work showing how post translational modifi cations
within the luminal domain of STIM1 aff ects the structural mechanisms of Ca2+ sensing. Ultimately, the post-translation
modifi cation driven STIM1 structural and biophysical changes have implications in the agonist induced hypertrophic response
and pinpoint a new therapeutic target for heart disease.

Toshiya Senda

High Energy Accelerator Research Organization (KEK), Japan

Title: Automated systems for X-ray crystallography at Photon Factory
Biography:

Toshiya Senda has completed his PhD from Nagaoka University of Technology, Niigata, Japan in 1995. He was a Research Associate in Nagaoka University of Technology
during 1995-2001 and a Senior Researcher in Institute of Advanced Industrial Science and Technology during 2001-2012. Now, he is the Director/Professor of Structural
Biology Research Center of High Energy Accelerator Research Organization (KEK) in Japan. He was awarded the CrSJ (Crystallographic Society of Japan) award in 2014.

Abstract:

X-ray crystallography has been the most widely utilized method for 3D structure determination of biological macromolecules
at atomic resolution. Since modern biology is studied at the molecular level, the 3D structures of biological macromolecules
provide critical information for rational biological studies. X-ray crystallography is therefore becoming an essential tool for
molecular and cellular biology and is increasingly being adopted by non-crystallographers. To facilitate this movement, the
Structural Biology Research Center (SBRC) in KEK has developed several robotics and automation systems for crystallization,
data collection, and structure determination. For the initial crystallization screening, we have developed protein crystallization
system (PXS) that can automatically perform initial crystallization screenings in a sitting drop setting. Th e PXS possesses
an automated image acquisition function for crystallization droplets. Th e obtained images can be checked from outside the
KEK campus via a VPN connection. Moreover, the crystallization plate can be directly mounted to a specifi c goniometer at
BL-17A for in situ data collection. Our protein crystallography beamlines, BL-1A, 5A, 17A, AR-NW12A, and AR-NE3A,
are highly automated by the PReMo program. Once a cassette containing protein crystals is set to the system, a robot can
mount a crystal to the goniometer in a beamline; do the centering of the crystal and start data collection. Beginning in May
2018, we will provide a fully automated data collection service for users. In this service, the obtained diff raction data sets will
be automatically processed by a data-processing routine in PReMo. Th e processed data can then further be analyzed with
PReMo for structural determination using the SAD method. Although at present the structure determination following the
data processing must be performed manually, we are developing a new routine for automated structure determination. It is
of note that all these automated functions are based on a database system in PReMo, and thus data from crystallization to
structure determination can be treated in an integrated manner. Our service is expected to provide the basis of an intelligent
system for crystallography.

Grace E Stutzmann

The Chicago Medical School - Rosalind Franklin University, USA

Title: Calcium signaling in the CNS in health and neurodegenerative disease
Biography:

Grace E Stutzmann focuses on early mechanisms of neurodegenerative disorders. She received her PhD from New York University, Center for Neural Science and
completed her Postdoctoral fellowships at Yale University and UC Irvine. She is currently an Associate Professor at Rosalind Franklin University/The Chicago Medical
School and serves as the Director for the Center for Neurodegenerative Disease and Therapeutics. Her research investigates effects of calcium signaling dysregulations
associated with AD on neuronal physiology, synaptic transmission, and histopathology. She uses transgenic mice expressing human mutations that cause familial AD and
employs techniques such as in vitro electrophysiology and 2-photon calcium imaging to study activity in neurons, in addition to immuno-based assays, molecular biology
and behavioral approaches. Her research has shown that specifi c calcium-mediated signaling pathways are dysregulated in AD and over time, facilitate amyloid plaques
and tangles formation, interfere with memory encoding circuits, and eventually can kill the cell.

Abstract:

Statement of the Problem: A shared aspect among many neurodegenerative disorders is a limited understanding of cellular
disease mechanisms. In our research, we have found that dysregulated calcium channels play a central and early role in driving
diseases such as Alzheimer’s, Huntington’s and brain injury. Specifi cally, defects in intracellular calcium stores within the
endoplasmic reticulum (ER) and their resident release channels such as the ryanodine receptor (RyR) are upstream of the
major disease features such as protein aggregates and synaptic defi cits. By focusing upon the proximal calcium channelopathy,
rather than later-stage features, signifi cant advances in understanding disease etiology may be made.
Methodology & Th eoretical Orientation: Whole cell patch clamp electrophysiology combined with 2-photon calcium
imaging are used to record signaling from hippocampal neurons in acute brain slices from mouse models of Alzheimer’s
disease (AD) and human neurons transformed from AD patients. Immunoassays and qRT PCR probe protein and transcript
levels in mouse and human neurons.
Findings: In early stage AD mice, RyR-evoked calcium signals are signifi cantly greater than non-transgenic controls, while
calcium signaling though voltage-gated channels and NMDA receptors are not diff erent. Similar observations were made
in human neurons from AD patients. Additionally, RyR 2 message is increased in both the mouse and human AD neurons.
Pharmacologically normalizing RyR-calcium release restores RyR 2 levels in mice, as well as reduces many of the AD features
including beta amyloid, hyperphosphorylated tau and synaptic loss.
Conclusion & Signifi cance: Dysregulated ER calcium channel functioning is an early feature of AD in mouse models and
human samples and likely initiates a pathogenic cascade in the proximal disease stages. Because of the multiple and essential
roles of calcium signaling in neurons, alterations in RyR-channel properties can generate multiple downstream maladaptive
eff ects such as those seen in neurodegenerative disorders.

Biography:

Jan B Parys is a faculty member since 1997, ordinary Professor of Physiology since 2006 and Head of the Laboratory of Molecular and Cellular Signaling at the KU
Leuven, Belgium since 2013. His research always concerned the understanding of the mechanisms for Ca2+ homeostasis in general and the structure, function and
regulation of the inositol 1, 4, 5-trisphosphate receptor (IP3R), a central player in the initiation and propagation of intracellular Ca2+ signals, in particular. More recently, he
focused on the role of the IP3R in conjunction with other Ca2+-transporting proteins of the endoplasmic reticulum, the mitochondria and the lysosomes in cell survival and
cell death, and especially in the processes of apoptosis and autophagy. Since 2010 he is Secretary-General of the European Calcium Society (ECS) and was organizer
or co-organizer of multiple international workshops (2013) and meetings (2008, 2014, and 2016) of the ECS.

Abstract:

The inositol 1, 4, 5-trisphosphate (IP3) receptors (IP3Rs) are ubiquitously expressed Ca2+ release channels of the endoplasmic
reticulum (ER). Th e resulting Ca2+ signals are instrumental in the regulation of many intracellular processes. It is now
becoming increasingly clear that contact sites between ER and mitochondria or between ER and lysosomes is very important
for controlling mitochondrial energetics, autophagy and apoptosis. Anti-apoptotic Bcl-2 leads to increased cell survival by both
blocking pro-apoptotic family members like Bax and Bak and by inhibiting IP3R mediated Ca2+ transfer to the mitochondria.
As Bcl-2 is upregulated in a variety of cancers, including diff use large B-cell lymphoma (DLBCL) and chronic lymphocytic
leukemia (CLL), we developed in collaboration with Clark W Distelhorst (Case Western Reserve University, USA) BIRD-2, a
peptide tool that disrupts Bcl-2/IP3R interaction by binding to the Bcl-2 BH4 domain, increasing IP3 induced Ca2+ release and
triggering apoptosis of DLBCL cells and of primary CLL cells. Interestingly, DLBCL cell lines with high expression levels of
the high affi nity IP3R2 isoform, were the most sensitive to BIRD-2. In addition to high levels of IP3R2 expression, constitutive
IP3 signaling downstream of the tonically active B-cell receptor increases sensitivity towards BIRD-2. Under standard growth
conditions, this constitutive IP3 signaling fulfi lled a pro-survival role, since inhibition of phospholipase C using 2.5 μM U73122
caused cell death in various DLBCL cell lines and primary CLL cells, but at lower concentrations protected against BIRD-
2-induced Ca2+ release and apoptosis. Taken together, our data indicate that on the one hand, constitutive IP3 signaling is
necessary for cancer cell survival and proliferation but that on the other hand to survive this chronic signaling the cancer cells
are dependent on high Bcl-2 levels to prevent excessive Ca2+ release via IP3Rs. Disrupting Bcl-2/IP3R interaction switches
constitutive IP3 signaling to a pro-death signal and may therefore lead to a novel therapeutic approach

  • Structural Enzymology | Computational Approaches | Recent Advancements in Structural Biology
Location: Spreewald
Biography:

Ray Unwalla is a computational chemist at Pfi zer, USA working in the Infl ammation and Immunology Department. He has several years of drug discovery experience
and has expertise in the area of nuclear hormone receptors, kinases and GPCRs. He has an in depth knowledge of various computational tools and techniques to
provide insightful design hypothesis and specifi c compound proposals that test design hypothesis. He was involved in project teams that successfully delivered two drug
candidates i.e., Dutasteride for benign prostate hyperplasia, Duavee for hormone replacement therapy and six clinical candidates for the progesterone, ERα LXRβ, SARM,
JAK1 and JAK3 targets.

Abstract:

The steroids testosterone and dihydrotestosterone (DHT) are androgens that play an important role in the development and
maintenance of a variety of physiological responses such as male sexual function, bone density, muscle mass and strength.
Th e androgen receptor is a nuclear hormone receptor that is expressed in many tissues and is responsible for mediating the
actions of testosterone and DHT. Patients that have defects in the androgen receptor or have androgen defi ciencies can be
eff ectively treated with exogenous testosterone and other steroidal androgens as a hormone replacement therapy. Th e anabolic
eff ects of testosterone have shown benefi t in age related decline of bone density and muscle mass. However, the side eff ect profi le
of testosterone and other currently available anabolic steroids precludes their wide spread use and the chronic administration
of steroidal androgens is associated with potential serious side eff ects such as hepatotoxicity, prostate hypertrophy and cancer.
In addition, the oral bioavailability of testosterone is poor and the route of its administration is generally through topical
formulations. We will describe our eff orts to fi nd novel series of oral tissue selective androgen receptor modulators (SARM)
i.e., cyanopyrroles and indolines that selectively promote muscle growth while showing reduced androgenic eff ects on the
prostate and seminal vesicles. Using a docking approach, we were able to delineate the binding mode of these series and further
optimize the potency. An x-ray structure of a lead compound 7 bound to AR ligand binding domain revealed an interesting
electrostatically unfavorable interaction of the 3-fl uorophenol group with a carbonyl backbone of Leu704 residue. QM based
intermolecular potential calculation performed to understand the strength of this interaction along with ligand strain energy
calculations indicate a small energy penalty of ~0.6 kcal/mol paid by the ligand to adopt the bound state conformation

Biography:

R Holland Cheng is a Professor of Molecular and Cellular Biology at University of California. He received a Master degree in 1989 and a PhD degree in 1992 from Purdue
University. He has served as a Panel Reviewer for NIH programmed project grant, NIH Microbiology & Infectious Diseases Research Committee, NIH National Center
for Research Resources, Wellcome Trust, UK and Earth & Life Sciences Council, Netherlands (2002). He has actively served on the advisory board of the International
Microscopy Congress plus regional microscopy societies, American Biographical Institute, and International Virus Assembly Symposium. He is also a regular member of
the American Chemical Society, American Society for Microbiology and American Biophysical Society. He has published nearly fi fty papers in peer reviewed journals and
his research mainly focuses on proteome imaging of macromolecular systems.

Abstract:

Cryogenic electron microscopy (cryo-EM) provides a visible and quantitative approach to investigate the function of
structural proteins of viruses, both in assembly and disassembly. Th e fundamental biochemical properties of metastable
intermediates and stable mature form of viral capsids, responsible for their capacity to resist extreme pH and digestive enzyme
degradation, are revealed through cryo-EM. In this presentation, the structural functionality of Hepatitis E viral nanoparticles
(HEVNP), as a multimodal delivery platform, is assessed for the delivery of medicals or agents to target tumors through
both circulation and mucosal routes. Aided by multimodal imaging and integrative machine learning networks, the targeting,
tissue distribution, and payload packaging are investigated and demonstrated. Together, cryo-EM and cellular tomography are
exemplifi ed in the recent outcomes of unveiling the molecular mechanisms essential for platform vector design towards the
success of constructing a non-invasive, mucosal targeting system.

Biography:

Abstract:

A large number of viruses encode small proteins which are expressed in the infected cell to form ion and small molecules
conducting channels. Th ese proteins are also called viroporins. For some of the viruses these proteins are essential for
infectivity cycle and thus, they comprise interesting drug targets. From the structural and mechanistic point of view, they
comprise interesting tools to study membrane protein assembly and dynamics in particular when the outcome of the assembly
is an ion conducting system. Computational tools play an important part in this investigation. Using a 2D docking tool
structural models of polytopic membrane proteins such as p7 of hepatitis C virus (HCV) with two transmembrane domains
(TMDs) and E5 of human papilloma virus – 16 (HPV-16) with three TMDs are generated in a putative functional form.
From molecular dynamics (MD) simulations of these proteins embedded in a hydrated lipid bilayer ion selectivity of these
proteins is investigated also by calculating potential of mean force (PMF) profi les of ions along the conducting pathway.
Th e calculations support weak selectivity of these channel proteins, strengthening the quality of the modeled structures. In
addition, full correlation analysis (FCA) provides insight into the dynamic of these proteins. Th e individual subunits of the
protein assemblies show asymmetric behavior during equilibrium dynamics, allowing for proposing several models of gating.

Biography:

Liliane Mouawad was always interested in understanding the mechanism of action of proteins or protein assemblies. This understanding may be based on either molecular
simulations or on experiments like NMR. But her expertise is primarily in molecular dynamics simulations and more precisely in normal mode analysis (NMA). She
has developed several methods going from the calculation of normal modes of very large systems or of images, to the calculation of the pathway between two protein
conformations or to the prediction of the compactness of a calcium-binding protein. Recently, she was also involved in docking and virtual screening themes, where she
has acquired enough expertise to develop a new consensus methodology to overcome some issues observed in these approaches.

Abstract:

The Ca2+ pump SERCA1a is a P-type ATPase, localized in the sarcoplasmic reticulum membrane of striated muscle cells.
SERCA1a is involved in the contraction/relaxation process by fast pumping the cytoplasmic Ca2+ into the reticulum.
Th roughout its catalytic cycle, SERCA1a presents two major conformations: the E1 conformation where its Ca2+ channel is open
toward the cytoplasm and the E2 conformation where this channel is open toward the lumen. Th is conformational transition
is enhanced by ATP phosphorylation which occurs aft er Ca2+ binding by SERCA1a. Sarcolipin (SLN), a transmembrane helix
of 31 residues regulates SERCA1a by diminishing its affi nity to Ca2+. Th e mechanism of this regulation is not elucidated yet
despite the knowledge of the crystal structure (see fi gure). To decipher this mechanism, we performed normal mode analysis
(NMA), in the all-atom model on two systems, in the presence and the absence of SLN in a POPC membrane and one layer of
water for the soluble parts of the protein. Th is analysis showed that both systems are prone to go toward the conformation of
2Ca2+ E1 more easily than toward that of E2. However, both transitions seem more diffi cult in the presence of SLN, because of
some specifi c interactions with SERCA1a that result in additional fl uctuations of SERCA1a-SLN. A long-distance transmission
of information (over 35 Å) within the protein was also observed, explaining the phosphorylation diffi culty in the complex.
Th ese results provide new insights into the mechanism of the SERCA1a enzymatic cycle and its regulation by SLN.

Biography:

Sangwook Wu received his BA degree in Biochemistry from Yonsei University, Korea in 1990. After working as a Scientist at Samsung Display Devices during 1995-1999,
he obtained his PhD in Theoretical Condensed Matter Physics from Iowa State University during 1999-2005. He joined the Computational Biophysics Lab at UNC-Chapel
Hill (Dr. Lee Pedersen) as postdoctoral research associate during 2005-2014. From 2014 to the present, he has been a faculty member at Pukyong National University in
Korea. His research interests are in the areas of computational dynamics of biological macromolecules.

Abstract:

Non-receptor tyrosine kinase c-Src plays a critical role in numerous cellular signaling pathways. Activation of c-Src
involves a change from its inactive to the active state accompanied by large-scale conformational change depending
on the phosphorylation state of two major phosphorylation sites, Tyr416 and Tyr527. A detailed mechanism for the entire
conformational transition of c-Src via phosphorylation control of Tyr416 and Tyr527 is still elusive. In this study, we investigated
the inactive to active conformational change of c-Src by targeted molecular dynamics simulation. Based on the simulation, we
proposed a dynamical scenario for the activation process of c-Src. A detailed study of the conformational transition pathway
based on network analysis suggests that Lys321 plays a key role in the c-Src activation process.