The Research Center for Molecular Mechanism of Aging and Age-related Diseases, Landau Phystech-School of Physics & Research of the Moscow Institute of Physics and Technology (MIPT) invites you to take part in the International Seminar "Molecular mechanisms of aging and age-related diseases", which will be held on October 2 - 30, 2019 at MIPT campus (Dolgoprudny).
The main purpose of the International Seminar is to discuss key achievements in various research fields of cellular and molecular biophysics. Advanced biophysics methods, including modern fluorescence microscopy and CryoEM, X-ray crystallography and their application to the most pressing scientific challenges, such as molecular mechanisms of aging, cancer, neurodegenerative, cardiovascular and infectious diseases are taking the center stage on the International seminar agenda. The program includes public lectures and round tables.
Open Lecture Schedule
- October 7, 2019
- October 16, 2019
- October 18, 2019
- October 28, 2019
- October 29, 2019
- October 30, 2019
107 PhystechBIO Bld.
Prof. Dr. Dieter Willbold / Forschungszentrum Jülich, Heinrich-Heine-Universität Düsseldorf, Germany
"Direct elimination of Aβ oligomers by the orally available clinical stage anti-prionic compound PRI-002 reverses cognition deficits and decelerates neurodegeneration in Alzheimer’s disease animal models"
The mechanisms defining the function and dysfunction of the proteins involved in neurodegenerative and infectious diseases — especially Alzheimer’s — are fairly well studied. Based on the available information, Dieter Willbold’s research group quantitatively investigated the structures of these proteins and the ligand interactions with them at a high spatial resolution. The results of this research helped develop a clinical strategy, as well as new therapeutic and diagnostic approaches targeting amyloid beta oligomers.
107 PhystechBIO Bld.
Dr. Richard Lozier (Independent editor of scientific manuscripts)
"44 years ago: the first photocycle model of bacteriorhodopsin"
Dr. Igor Chizhov / Institute for Biophysical Chemistry, Hannover Medical School, Germany
"How many models of photocycles exist now and how we can live with this"
Streaming Lecture Hall, PhystechCifra Bld.
Riccardo Rosselli / NIOZ- The Netherlands Institute for Sea Research, The Netherlands
"Novel approaches and metagenomic developments to analyze environmental rhodopsin diversity"
Microbial rhodopsins are encoded by Prokaryotes, unicellular Eukaryotes and viruses and represent the evolutive response of heterotrophs to the natural light utilization. The advent of culture-independent DNA sequencing approaches has shed novel insights into the astonishing variety that characterizes microbial genomes. The possibility to analyze nucleic acids directly extracted from the environment, encouraged global sampling expeditions (e.g., TARA-Oceans, Malaspina, OSD-consortium) aimed at exploring genomic composition and diversity of entire ecosystems, and providing public available data for the scientific community.
This data definitively showed that microbial rhodopsins previously known from culture collections barely represent the diversity that the group has in oceans and other aquatic environments. Current bioinformatic approaches to analyze environmental gene diversities, are largely based on sequence similarity searches. They allowed to retrieve novel rhodopsin groups and rhodopsins with novel functions. Nevertheless, in order to expand the knowledge about this protein family, the research for environmental rhodopsins should upgrade algorithms and develop approaches that might allow to investigate also on clades which sequences remarkably diverge from those which are already known, as the novel group of microbial rhodopsins of type-3. To achieve this objective, a bioinformatic platform is is being set that might spot divergent rhodopsins and proteins that for the moment represent the dark side of the natural light utilization.
107 PhystechBIO Bld.
Alexander S. Mishin and Mikhail S. Baranov / Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russia
"Fluorogens for live-cell imaging and nanoscopy"
Fluorogenic dyes are non-fluorescent in solution but become highly-emissive in complex with the target molecule. We exploit this principle in the rational design of bipartite protein labeling systems, tailored for live-cell imaging and super-resolution microscopy. In this talk, we will report on current progress in the development of fluorogen-activating proteins derived from either bacterial lipocalin Blc or photoactive yellow protein. We will also cover the fundamentals of the environmental sensitivity of a unique group of synthetic derivatives of natural fluorogen - chromophore of the Green Fluorescent Protein. In conclusion, we will review the advantages of fluorogenic dyes and fluorogen-activating proteins for live-cell imaging and nanoscopy.
107 PhystechBIO Bld.
Andrei Gilep / Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Belarus
"Interactomics approaches in study of functions and molecular organisation of protein complexes"
Our approach to study molecular organisation and functions of protein complexes involves the use of a combined interactomics approach consisting of biospecific oriented immobilization of protein of interest, affinity isolation of target molecules from a lysate of biological material based on a specific interaction, phage-display technologies and validation of protein complexes by biosensors using recombinant purified proteins. Biospecific oriented immobilization is carried out by fusion of recombinant protein with Avi-tag for in vivo/in vitro site-specific biotinylation and specific interaction with streptavidin-bearing sorbent. The interaction of an immobilized protein (“bait protein”) and functionally active potential partner proteins are then analyzed using LC-MS-MS. In addition to methods based on the affinity isolation of target molecules, we use methods of the reverse interactomics, which consist of using peptide phage display methods to determine specific peptide motifs interacting with target membrane proteins. An analysis of the pool of such sequences obtained after enrichment of the phage library for “bait protein” allows us to assume the protein partners of this protein, which includes these sequences. A high-performance peptide epitope scan of the sequence of putative protein partners can determine the interface of interaction with the “bait protein”. Detected PPIs are then validated using biosensor methods (SPR - surface plasmon resonance, BLI - biolayer interferometry). We also use phage display technology to generate recombinant antibodies. Single domain antibodies to membrane proteins could serve as a tool for their trapping in a specific conformational state, facilitating structural studies and drug discovery. As a tool for single-domain antibodies rapid and reliable generation, we can provide synthetic VHH library with a diversity of 1e10 clones. An optimized high-throughput screening protocol allows generation of antibody binders in just two weeks. We apply interactomics approaches for study of proteins involved in biosynthesis of steroids, eicosanoids and porphyrins.
Andrei Gilep / Institute of Bioorganic Chemistry, National Academy of Sciences of Belarus, Belarus
"Human cytochrome P450 as drug target for anticancer chemotherapy"
Cytochrome P450 (CYP) is a superfamily of hemeproteins involved in oxidative transformation of endogenous compounds (such as cholesterol, vitamin D3, eicosanoids etc.) and xenobiotics including drugs. Essential role of CYP enzymes in steroid biosynthesis make some of them validated drug targets. Azole antifungal drugs currently in use were discovered empirically before the target, sterol 14α-demethylase (CYP51) was identified. CYP51 is evolutionary conserved enzyme with preserved function and hence the selectivity could be achieved by exploiting differences in enzyme structure between host and pathogen. However, human CYP51 could be considered as target itself to reduce cholesterol. We solved human CYP51 crystal structures, alone and in complex with azole drugs, as a template for rational drug design. Based on accumulated knowledge and data obtained by us and others we are going to apply machine learning to identify new scaffolds for selective inhibition.
Dr. Albert Guskov / The University of Groningen, The Netherlands
"The ABC transporter Rv1819c from Mycobacterium tuberculosis mediates uptake of unrelated hydrophilic compounds"
The Mycobacterium tuberculosis (Mtb) ATP-binding cassette (ABC) transporter Rv1819c is implicated in tuberculosis (TB) pathogenesis. Here, I will present a structure of Rv1819c, and show that while Rv1819c exhibits the ABC exporter fold, the protein mediates import of the unrelated, hydrophilic compounds vitamin B12 (cobalamin) and bleomycin. The poorly specific importer activity can be explained by the presence of an enormous, membrane- spanning, and water-filled space (of ~7,700 Å3) in the protein interior, which is lined with a hydrophilic surface. In the determined structure, this protein cavity is occluded, and comparison with other ABC transporters suggests that it can open alternatingly to either side of the membrane. Lack of coupling between ATPase activity and presence of transported substrate, suggests that the protein provides Mtb with basal multi-solute transport capacity.
Sergey Permyakov / Institute for Biological Instrumentation of the Russian Academy of Sciences, Russia
"Interactions between cytokines and S100 proteins: next complexity level in regulation of biological processes"
Cytokines is a family of over hundred small extracellular signaling proteins mediating cellular communication in virtually all biological processes. Cytokines are engaged into progression of immune, inflammatory, oncological and reproductive system diseases, enabling use of cytokines, inhibitors of cytokines or their receptors in therapy of numerous diseases. S100 proteins is a family of over 24 small metal-binding intra- and extracellular regulatory proteins of vertebrates, involved into various physiological processes. Some S100 proteins are associated with heart, immune, inflammatory, oncological and neurodegenerative diseases. Some of the S100 proteins are secreted or released into extracellular space upon cell damage and bind to certain receptors, thereby affecting intracellular signaling. The same effect is achieved via interaction of the S100 proteins with ligands of such receptors as EGFR. Several extracellular S100 proteins belong to the families of alarmins or damage associated molecular patterns. The latter are related to progression of inflammation, cancer, and ageing. While effects exerted by individual cytokines and extracellular S100 proteins on some cellular processes have been studied in detail, direct interactions between cytokines and S100 proteins remain unexplored with regard to incidence of this phenomenon and its functional significance. Here we’ll review the progress in search and validation of interactions between cytokines and extracellular S100 proteins. The discovery of novel S100-cytokine interactions contributes to recognition of this phenomenon as an additional complexity level in regulation of biological processes mediated by these proteins, thereby providing a basis for development of next-generation drugs.
Norber Dencher / Darmstadt University of Technology, Germany; Moscow Institute of Physics and Technology, Russia
"Interaction of Alzheimer’s disease triggering amyloid beta peptides with membranes and organelles: bioenergetical consequences"
Alzheimer’s disease affects 47 million people worldwide. No significant progress has been made in research for early diagnosis and efficient therapy. One of the characteristic histopathological markers of AD is the presence of extracellular plaques consisting predominantly of amyloid-β peptides (Aβ) with 40 or 42 amino acids. Recent research suggests that monomers and/or soluble oligomers of Aβ are responsible for AD symptoms but not the extracellular plaques. To study cellular and organelle trafficking of Aß, to identify its target(s), Aß42 peptide monomers/small oligomers were externally applied to mammalian cells (human neuroblastoma and rat oligodendroglia). Monomeric/oligomeric peptides entered cells, as proven by confocal fluorescence microscopy by employing fluorescently labeled Aβ42 peptides. In time and space the pathway of Aß peptides from the outside of the cell across the plasma membrane to internal target membranes of specific organelles was tracked. Aβ peptides initially co-localized with the plasma membrane and thereafter entered the cells and trafficked to organelles, e.g., predominantly to lysosomes. The deep insertion of Aß in lipid bilayers that subsequently induced membrane perturbations and alterations in lipid dynamics was verified by neutron scattering and fluorescence polarization. Aß affected numerous physiological cell parameters, such as ROS and ATP concentration, viability (necrosis/apoptosis), mitochondrial membrane potential and viscosity. The observed Aß-induced alterations might cause neuronal cell death and AD pathology. Furthermore, our data challenge the “Aß extracellular plaque hypothesis of AD”.
107 PhystechBIO Bld.
Fonin Alexander / Institute of Cytology Russian Academy of Sciences, Russia
"Mechanism of formation and function of stress-induced membraneless organelles"
In eukaryotic cell there are a large number of functional structures which are not limited by the membrane, called membraneless organelles. For a long time, each such structure was studied by cell biologists independently of other similar structures. Only at the beginning of the 21st century physicists and molecular biologists paid attention to the fact that the formation of all membraneless organelles is based on a single principle, namely phase separation of biopolymers in macromolecular crowding conditions and that intrinsically disordered proteins play a key role in this process. Thus, it turned out that membraneless organelles must be studied from a single point of view. Currently, the number of works devoted to the study of these objects is growing avalanche- like. This is primarily caused the fact that, due to their structure dynamics, these organelles play a significant role in the spatial-temporal organization of intracellular space. A slight distortion of the external or internal conditions may cause their disassembly, or, conversely, the transition of membraneless organelles into a gel-like state arising from irreversible aggregation of their protein constituents. The highly dynamic structure and composition, the multi-component content and poly-functionality of membrane-less organelles allow them to ensure finely tuned regulation of various intracellular processes. In this regard, even slight disturbance of the activity of membrane-less organelles can lead to the dysregulation of the intracellular pathways. We intend to focus on the factors affecting the functional activity and formation of membraneless organelles. The objects of our investigations were PML bodies and artificial light-induced stress- granules, which properties are strongly depended on external stimulus. We have established that the molecular mechanisms of the formation and functioning of nuclear PML bodies (PML-NB) associated with the process of Alternative Lengthening of Telomeres (ALT-associated PML-NB, APB) are dependent on the polyampholyte blocks in the disordered C-terminal regions of PML isoforms. This protein is the scaffold protein of PML-bodies. Also, we have shown that these polyampholyte blocks are important for light-induced formation of artificial stress granules.
Vedunova Maria, Babaev Alexey / Lobachevsky University, Russia
"Brain-derived neurotrophic factor preserves the functional activity of neural networks in the β-amyloidopathy model in vitro"
Alzheimer's disease (AD) studies are becoming more relevant year by year. The reason of such importance associated with an increase in life expectancy of the population and accumulation of information about AD polyethyology. Alzheimer’s disease (AD) is a widespread chronic neurodegenerative pathology characterized by synaptic dysfunction, partial death of neurons, cognitive and memory impairments etc. It is currently considered that the major hallmarks of AD are the presence of extracellular senile amyloid plaques formed by various types of amyloid proteins (Aβ), and the formation and accumulation of intracellular neurofibrillary tangles. However, there is still no effective therapeutic method for AD correction and relevant experimental models for studying the changes in neural network activity, the features of intercellular signaling and drugs effects on the functional activity of nervous cells during AD development. In this work, we have created experimental model of amyloidopathy using primary hippocampal cultures. The model is based on chronic application of amyloid beta 1-42 (Aβ1-42). Chronic application of Aβ1-42 allows quickly simulation of significant neurodegenerative changes in the primary hippocampal cultures, leading to sharp impairments in neural network calcium activity and increase cell death. Using this model and multielectrode arrays, we studied the influence of amyloidopathy on spontaneous bioelectrical neural network activity. We showed that the formation of amyloid conglomerates decreased the number of network bursts and spikes in a burst and disrupted the spatial network structure manifested in a decrease of the number of key network elements (hubs) and the number of connections between the network elements. Our study also revealed that application of BDNF in the form of recombinant protein and in a case of its hyperexpression by an adeno-associated virus vector partially prevents amyloidopathy-induced neurodegenerative phenomena. The use of BDNF maintains cell viability and spontaneous bioelectrical and calcium network activity in primary hippocampal cultures. The internal functional structure of neural networks, the number of hubs and connections between active elements in the network were partially preserved.
Dr. Ravinder Anand-Ivell / University of Nottingham, UK
"Gonadal physiology as a key to lifelong health and morbidity"
In the fetus the gonads are one of the earliest functional organ systems to develop and produce hormones which influence the growth and development of all other systems, thereby providing an endocrine blueprint with effects evident across the lifespan and into old age. Whilst the sex steroids are the best known of these hormones, the gonads also produce what are called ‘neohormones’ which have evolved to address the specialist physiology of the mammal, such as viviparity, internal fertilisation, pregnancy and lactation. The insulin-like peptides relaxin and INSL3 are best known of these hormones and act through large GPCRs, called RXFP1 and RXFP2, respectively. These hormone-receptor systems are active across the reproductive lifespan and decline in old age in both males and females. Besides their involvement in reproduction, recent research is showing how they also influence other organs, such as the cardiovascular, musculo-skeletal, and renal systems, implying that the gonadal production of these hormones can also affect general health and morbidity.