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Author: Brandon Mac Butler
Publisher:
ISBN:
Category : DNA-protein interactions
Languages : en
Pages : 135
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Book Description
Proteins are essential for most biological processes that constitute life. The function of a protein is encoded within its 3D folded structure, which is determined by its sequence of amino acids. A variation of a single nucleotide in the DNA during transcription (nSNV) can alter the amino acid sequence (i.e., a mutation in the protein sequence), which can adversely impact protein function and sometimes cause disease. These mutations are the most prevalent form of variations in humans, and each individual genome harbors tens of thousands of nSNVs that can be benign (neutral) or lead to disease. The primary way to assess the impact of nSNVs on function is through evolutionary approaches based on positional amino acid conservation. These approaches are largely inadequate in the regime where positions evolve at a fast rate. We developed a method called dynamic flexibility index (DFI) that measures site-specific conformational dynamics of a protein, which is paramount in exploring mechanisms of the impact of nSNVs on function. In this thesis, we demonstrate that DFI can distinguish the disease-associated and neutral nSNVs, particularly for fast evolving positions where evolutionary approaches lack predictive power. We also describe an additional dynamics-based metric, dynamic coupling index (DCI), which measures the dynamic allosteric residue coupling of distal sites on the protein with the functionally critical (i.e., active) sites. Through DCI, we analyzed 200 disease mutations of a specific enzyme called GCase, and a proteome-wide analysis of 75 human enzymes containing 323 neutral and 362 disease mutations. In both cases we observed that sites with high dynamic allosteric residue coupling with the functional sites (i.e., DARC spots) have an increased susceptibility to harboring disease nSNVs. Overall, our comprehensive proteome-wide analysis suggests that incorporating these novel position-specific conformational dynamics based metrics into genomics can complement current approaches to increase the accuracy of diagnosing disease nSNVs. Furthermore, they provide mechanistic insights about disease development. Lastly, we introduce a new, purely sequence-based model that can estimate the dynamics profile of a protein by only utilizing coevolution information, eliminating the requirement of the 3D structure for determining dynamics.
Author: Brandon Mac Butler
Publisher:
ISBN:
Category : DNA-protein interactions
Languages : en
Pages : 135
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Book Description
Proteins are essential for most biological processes that constitute life. The function of a protein is encoded within its 3D folded structure, which is determined by its sequence of amino acids. A variation of a single nucleotide in the DNA during transcription (nSNV) can alter the amino acid sequence (i.e., a mutation in the protein sequence), which can adversely impact protein function and sometimes cause disease. These mutations are the most prevalent form of variations in humans, and each individual genome harbors tens of thousands of nSNVs that can be benign (neutral) or lead to disease. The primary way to assess the impact of nSNVs on function is through evolutionary approaches based on positional amino acid conservation. These approaches are largely inadequate in the regime where positions evolve at a fast rate. We developed a method called dynamic flexibility index (DFI) that measures site-specific conformational dynamics of a protein, which is paramount in exploring mechanisms of the impact of nSNVs on function. In this thesis, we demonstrate that DFI can distinguish the disease-associated and neutral nSNVs, particularly for fast evolving positions where evolutionary approaches lack predictive power. We also describe an additional dynamics-based metric, dynamic coupling index (DCI), which measures the dynamic allosteric residue coupling of distal sites on the protein with the functionally critical (i.e., active) sites. Through DCI, we analyzed 200 disease mutations of a specific enzyme called GCase, and a proteome-wide analysis of 75 human enzymes containing 323 neutral and 362 disease mutations. In both cases we observed that sites with high dynamic allosteric residue coupling with the functional sites (i.e., DARC spots) have an increased susceptibility to harboring disease nSNVs. Overall, our comprehensive proteome-wide analysis suggests that incorporating these novel position-specific conformational dynamics based metrics into genomics can complement current approaches to increase the accuracy of diagnosing disease nSNVs. Furthermore, they provide mechanistic insights about disease development. Lastly, we introduce a new, purely sequence-based model that can estimate the dynamics profile of a protein by only utilizing coevolution information, eliminating the requirement of the 3D structure for determining dynamics.
Author: Ke-li Han
Publisher: Springer Science & Business Media
ISBN: 3319029703
Category : Medical
Languages : en
Pages : 488
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Book Description
This book discusses how biological molecules exert their function and regulate biological processes, with a clear focus on how conformational dynamics of proteins are critical in this respect. In the last decade, the advancements in computational biology, nuclear magnetic resonance including paramagnetic relaxation enhancement, and fluorescence-based ensemble/single-molecule techniques have shown that biological molecules (proteins, DNAs and RNAs) fluctuate under equilibrium conditions. The conformational and energetic spaces that these fluctuations explore likely contain active conformations that are critical for their function. More interestingly, these fluctuations can respond actively to external cues, which introduces layers of tight regulation on the biological processes that they dictate. A growing number of studies have suggested that conformational dynamics of proteins govern their role in regulating biological functions, examples of this regulation can be found in signal transduction, molecular recognition, apoptosis, protein / ion / other molecules translocation and gene expression. On the experimental side, the technical advances have offered deep insights into the conformational motions of a number of proteins. These studies greatly enrich our knowledge of the interplay between structure and function. On the theoretical side, novel approaches and detailed computational simulations have provided powerful tools in the study of enzyme catalysis, protein / drug design, protein / ion / other molecule translocation and protein folding/aggregation, to name but a few. This work contains detailed information, not only on the conformational motions of biological systems, but also on the potential governing forces of conformational dynamics (transient interactions, chemical and physical origins, thermodynamic properties). New developments in computational simulations will greatly enhance our understanding of how these molecules function in various biological events.
Author: P. Jolles
Publisher: Birkhäuser
ISBN: 3034884583
Category : Science
Languages : en
Pages : 240
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Book Description
A wealth of information has accumulated over the last few years on the human genome. The new insights have completely changed the focus of protein analysis. It is no longer time-consuming analysis of unknown products, but rather selective identifications of individual forms, modifications and processings, and overall analysis of global protein outputs from cells and tissues in health and disease. This book gears to the rising need of sensitive, accurate, and fast separation and identification techniques in proteomics. It discusses current methodologies of modern protein analysis, from isolation and sample preparation, over analysis and identification, to final characterization. Several evaluations concentrate on the now productive approaches of two-dimensional gel electrophoresis and mass spectrometry, but alternative methods and further perspectives are also outlined. The book includes an overlook over current databases to connect protein analysis data with all available information,...
Author: Philip W. Bransford
Publisher:
ISBN:
Category :
Languages : en
Pages : 145
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Book Description
The volume of protein structure data has grown rapidly over the past 30 years, leaving a wake of facts that still require explanation. We endeavored to answer a few open questions on the structure-function relationship of intriguing mechanochemical protein systems. To this end this thesis work contains five studies that offer novel insights into molecular biomechanical systems that may guide future basic research or applications development. The first study concerns the biophysics of cadherin-mediated cell sorting observed in developing solid tissue. We investigated the evolutionary dynamics of the cadherin superfamily of cell-cell adhesion proteins to infer a structural basis for their paradoxical mixture of pairwise binding specificity and promiscuity. Our analysis predicts a small set of specificity-determining residues located within the protomer-protomer binding interface. The putative specificity-determinants form a design space with potential for engineering novel cell-cell adhesive interactions. The second study addresses the open question of how to automatically identify regions within a protein that engage in allosteric communication. To identify allostery we developed and tested two computational tools that operate on protein conformational dynamics data. These tools are useful for generating testable hypotheses about proteins with multiple functional sites for the design of non-competitive protein inhibitors. The third study asks, "What is the consequence of allosteric cooperation between the tandem binding sites in a class of proteins that bundle filamentous actin (F-actin)?" Through simulation we demonstrate that cooperative F-actin bundling tends to strengthen bundles by driving the formation of cross-links between neighboring filaments while depleting F-actin binding sites that are occupied but not cross-linked. We hence propose that allostery may be a natural feature of ABPs with tandem F-actin binding sites if nature indeed selects for sturdy F-actin bundles. The final two studies examine the impact of two structural perturbations to Factin on its mechanics. Using structure-based computer modeling we develop a simple explanation for the mechanism by which the structure of actin's polymorphic subdomain 2 mediates 4-fold changes in F-actin's flexibility. We further demonstrate that two calponin homology domains stabilize F-actin by binding in a configuration that tends to relax the stress concentration at actin-actin interfaces.
Author: Ivan Y. Torshin
Publisher: Nova Publishers
ISBN: 9781600210488
Category : Medical
Languages : en
Pages : 282
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Book Description
Biomedicine is one of the most important fields for the prospective applications of the information from human genome studies. However, there are many 'white spots' in the present-day understanding of the biomedical implications of this information. Given that at least half of the proteins in the established sequence of the human genome have no annotation whatsoever and that the sequence similarity searches are not likely to produce any, definite research strategies to analyse the functions of these unknown proteins as well as other enigmatic aspects of the human genome are being elaborated. The elaboration of the logistics of these research strategies, of the relevant computational methodologies as well as the general management of the informational complexity of the biological systems belong to the main tasks for the post-genomic bioinformatics. This volume concentrates on the role of the biophysical studies and biophysical concepts that can assist the endeavour.
Author: Dev Bukhsh Singh
Publisher: Academic Press
ISBN: 9780443219801
Category : Computers
Languages : en
Pages : 0
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Book Description
Genome Analysis: Principles and Methods provides recent and advanced information about genome analysis approaches and techniques to study and annotate the structure and function of the genome. It is a compendium of important topics such as NGS analysis, genome fragmentation and assembly, metagenomics, cloning and expression, physical marker analysis, transcriptome data analysis, sequence alignment and comparison, evolutionary analysis, SNP analysis, genome-based disease diagnosis and therapies, micro-RNAs, pharmacogenomics, genetic approaches to disease intervention, and challenges with opportunities in genome analysis and genomics, etc. The latest developments in the field are discussed, and key concepts introduced to ensure readers understand advanced concepts and methodologies in the area. The book serves as a valuable guide to the present, emerging, and evolving research methodologies in the field.
Author: Ferruccio Palazzesi
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
Author: National Research Council
Publisher: National Academies Press
ISBN: 0309184029
Category : Mathematics
Languages : en
Pages : 235
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Book Description
The Chemical Sciences Roundtable provides a forum for discussing chemically related issues affecting government, industry and government. The goal is to strengthen the chemical sciences by foster communication among all the important stakeholders. At a recent Roundtable meeting, information technology was identified as an issue of increasing importance to all sectors of the chemical enterprise. This book is the result of a workshop convened to explore this topic.
Author: Dennis R. Livesay
Publisher: Humana
ISBN: 9781493963072
Category : Science
Languages : en
Pages : 0
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Book Description
In Protein Dynamics: Methods and Protocols, expert researchers in the field detail both experimental and computational methods to interrogate molecular level fluctuations. Chapters detail best-practice recipes covering both experimental and computational techniques, reflecting modern protein research. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Protein Dynamics: Methods and Protocols describes the most common and powerful methods used to characterize protein dynamics.
Author: Isoken Omosefe Airen
Publisher:
ISBN:
Category :
Languages : en
Pages :
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Book Description
We set out to develop and apply a high-throughput cell-free protein synthesis (CFPS) platform that provides functional genomics information for a wide variety of open reading frames (ORFs). We then used this information to improve CFPS yields by 4- to 5-fold, depending on the protein product. With the increasing number of completed genome sequences and ongoing sequencing projects, the post-genomic era has ushered in the promise of complete understanding of biological systems. For such a task, the most important set of information is inarguably the knowledge of the function of each gene product. To lead this endeavor of discerning the properties and functions of the entirety of an organism's genes and gene products, the field of functional genomics has emerged. Functional genomics focuses on dynamic cellular aspects, such as gene transcription, translation, and protein-protein interactions, in attempts to understand the relationship between an organism's genome and its phenotype. Thus, the ultimate goal of such studies is to provide a more complete picture of how biological function arises from the hereditary information of a living system. However, despite this clear interest in analyzing the expression and function of gene products, the development of techniques to address the high-throughput needs of functional genomics has been challenging, given the large diversity of protein functions and physiochemical properties, such as molecular weight and hydrophobicity, as well as the varying expression levels of proteins within a cell. In light of these challenges, we developed a sequential CFPS platform, which is capable of characterizing a variety of diverse proteins in the context of the dynamic metabolic networks that exist in vivo. The first round of expression is directed by PCR-generated expression templates (ETs) and creates an array of cell extracts that are individually enriched with a single target gene product. This round of CFPS is terminated by the selective degradation of the linear DNA templates, and a subsequent round of protein expression is initiated by the addition of a plasmid ET for a reporter protein. The array is then screened to identify gene products that enhanced or inhibited the expression and folding of the reporter. With such a method, we expect that the observations will expand our knowledge of both cell-free and in vivo metabolism, as well as identify key factors and reactions that could potentially lead to improved in vitro transcription, translation and protein folding. CFPS systems offer attractive alternatives to conventional fermentation processes used for protein production. Although improvements in CFPS energetics and reaction conditions have greatly enhanced in vitro protein synthesis, we believe that there are still other issues limiting the productivity of the technology. For this reason, identifying targets that could further improve CFPS is desirable. To validate the developed sequential CFPS protocol, we conducted a genome-wide survey of the well-studied bacterium Escherichia coli (E. coli) to identify soluble gene products that influence the in vitro metabolism. With this method, we identified 139 gene products (79 positive and 60 negative effectors) that influenced the cell-free transcription, translation, and protein folding of our three reporter proteins, as well as the energy metabolism and RNA and protein stability in the CFPS system. Encouragingly, most of the observed effects were consistent with the accepted in vivo metabolic functions of the gene products. However, many were not and required subsequent assays and in-depth literature searches to suggest hypotheses for the in vitro activities of the identified gene products. In many cases, the observations illuminated principles and influences that are unknown, lesser known, or secondary functions that were not expected to influence the CFPS performance. The information from the genome-wide survey was then used to guide modifications of the CFPS system to improve the productivity and duration of in vitro protein synthesis, as well as the efficiency of protein folding. First, fifteen positive effectors were produced and supplemented into the expression reactions in various combinations of the effectors in order to identify cooperative interactions that further enhance system performance. Next, we constructed and evaluated four mutant E. coli strains with chromosomal deletions in non-essential genes that encode negative effectors identified by the genomic survey. We also re-optimized the small molecule metabolite environment in the CFPS reactions. Thus, in the improved in vitro expression system, energy generation, translation initiation and elongation, and protein folding were enhanced; the reaction pH was stabilized; the supplies of specific molecular substrates that are essential for protein synthesis were replenished; and mRNA transcripts were stabilized. With this new system, the total, soluble, and active yields of the several diverse proteins were enhanced by 300 to 400%. The functional genomic analysis of E. coli has greatly broadened our understanding of the biology of the organism. And with the use of species-independent translational leaders that can facilitate cell-free expression (Mureev et al., 2009), our sequential CFPS platform can be used for similar genome-wide surveys of most organisms. In this way, the vast wealth of information available in the sequenced genomes will be utilized, and our knowledge of these biological systems will be significantly improved. Furthermore, the forward, or targeted, metabolic engineering strategy that was used to enhance our CFPS system can be applied to the development and/or improvement of most organism-based in vitro protein expression platforms. These targeted metabolic changes will lead to more rapid and more significant enhancements than traditional improvement strategies, as well as bring us closer to a complete understanding of the biological systems.
