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Articles 1 - 30 of 84

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Ste5 Membrane Localization Allows Mapk Pathway Signaling In Trans Between Kinases On Separate Scaffold Molecules, Rachel E. Lamson, Matthew J. Winters, Peter M. Pryciak Jun 2019

Ste5 Membrane Localization Allows Mapk Pathway Signaling In Trans Between Kinases On Separate Scaffold Molecules, Rachel E. Lamson, Matthew J. Winters, Peter M. Pryciak

University of Massachusetts Medical School Faculty Publications

The MAP kinase cascade is a ubiquitous eukaryotic signaling module that can be controlled by a diverse group of scaffold proteins. In budding yeast, activation of the mating MAP kinase cascade involves regulated membrane recruitment of the archetypal scaffold protein Ste5. This event promotes activation of the first kinase, but it also enhances subsequent signal propagation through the remainder of the cascade. By studying this latter effect, we find that membrane recruitment promotes signaling in trans between kinases on separate Ste5 molecules. First, trans signaling requires all Ste5 domains that mediate membrane recruitment, including both protein-binding and membrane-binding domains. Second ...


Developing Tale Proteins As A Biosensor For Detecting Pathogen Specific Double-Stranded Dna, Kathrine Gaiko Apr 2019

Developing Tale Proteins As A Biosensor For Detecting Pathogen Specific Double-Stranded Dna, Kathrine Gaiko

Honors College Capstone Experience/Thesis Projects

Transcription activator-like effector (TALE) proteins are important for DNA binding. They bind to specific nucleotide sequences by the use of two residues in each repeat allowing them to target specific DNA sequences. Their modular structure makes TALEs advantageous over other DNA binding proteins such as zinc finger proteins. Zinc finger proteins (ZFP) use a finger like projection to bind 3-4 subsequence base pairs while TALE proteins use two amino acid residues in each repeat to bind one nucleotide. ZFP can use SEER-Lac system for colorimetric detection, while TALE proteins can use fluorescence labeling with Alexa for detection of binding to ...


Role Of Protein Charge Density On Hepatitis B Virus Capsid Formation, Xinyu Sun, Dong Li, Zhaoshuai Wang, Panchao Yin, Rundong Hu, Rundong Hu, Hui Li, Qiao Liu, Yunyi Gao, Baiping Ren, Jie Zheng, Yinan Wei, Tianbo Liu Apr 2018

Role Of Protein Charge Density On Hepatitis B Virus Capsid Formation, Xinyu Sun, Dong Li, Zhaoshuai Wang, Panchao Yin, Rundong Hu, Rundong Hu, Hui Li, Qiao Liu, Yunyi Gao, Baiping Ren, Jie Zheng, Yinan Wei, Tianbo Liu

Chemistry Faculty Publications

The role of electrostatic interactions in the viral capsid assembly process was studied by comparing the assembly process of a truncated hepatitis B virus capsid protein Cp149 with its mutant protein D2N/D4N, which has the same conformational structure but four fewer charges per dimer. The capsid protein self-assembly was investigated under a wide range of protein surface charge densities by changing the protein concentration, buffer pH, and solution ionic strength. Lowering the protein charge density favored the capsid formation. However, lowering charge beyond a certain point resulted in capsid aggregation and precipitation. Interestingly, both the wild-type and D2N/D4N ...


The Trim-Nhl Protein Nhl-2 Is A Novel Co-Factor Of The Csr-1 And Hrde-1 22g-Rna Pathways, Peter R. Boag, Gregory M. Davis, Shikui Tu, Rhys N. Colson, Joshua W. T. Anderson, Menachem J. Gunzburg, Michelle A. Francisco, Debashish Ray, Tuhin Maity, Monica Z. Wu, Quaid D. Morris, Timothy R. Hughes, Jacqueline A. Wilce, University Of Toronto, Zhiping Weng Feb 2018

The Trim-Nhl Protein Nhl-2 Is A Novel Co-Factor Of The Csr-1 And Hrde-1 22g-Rna Pathways, Peter R. Boag, Gregory M. Davis, Shikui Tu, Rhys N. Colson, Joshua W. T. Anderson, Menachem J. Gunzburg, Michelle A. Francisco, Debashish Ray, Tuhin Maity, Monica Z. Wu, Quaid D. Morris, Timothy R. Hughes, Jacqueline A. Wilce, University Of Toronto, Zhiping Weng

University of Massachusetts Medical School Faculty Publications

Proper regulation of germline gene expression is essential for fertility and maintaining species integrity. In the C. elegans germline, a diverse repertoire of regulatory pathways promote the expression of endogenous germline genes and limit the expression of deleterious transcripts to maintain genome homeostasis. Here we show that the conserved TRIM-NHL protein, NHL-2, plays an essential role in the C. elegans germline, modulating germline chromatin and meiotic chromosome organization. We uncover a role for NHL-2 as a co-factor in both positively (CSR-1) and negatively (HRDE-1) acting germline 22G-small RNA pathways and the somatic nuclear RNAi pathway. Furthermore, we demonstrate that NHL-2 ...


Pervasive Contingency And Entrenchment In A Billion Years Of Hsp90 Evolution, Tyler N. Starr, Julia Flynn, Parul Mishra, Daniel N. Bolon, Joseph W. Thornton Jan 2018

Pervasive Contingency And Entrenchment In A Billion Years Of Hsp90 Evolution, Tyler N. Starr, Julia Flynn, Parul Mishra, Daniel N. Bolon, Joseph W. Thornton

University of Massachusetts Medical School Faculty Publications

Interactions among mutations within a protein have the potential to make molecular evolution contingent and irreversible, but the extent to which epistasis actually shaped historical evolutionary trajectories is unclear. We addressed this question by identifying all amino acid substitutions that occurred during the billion-year evolutionary history of the heat shock protein 90 (Hsp90) ATPase domain beginning from a deep eukaryotic ancestor to modern Saccharomyces cerevisiae and then precisely measuring their fitness effects when introduced into both extant and reconstructed ancestral Hsp90 proteins. We find a pervasive influence of epistasis: of 98 derived states that evolved during history, most were deleterious ...


2-D Electrophoresis Modeling Of Multienzyme Cutting Of Polypeptides, Howard Mayes Oct 2017

2-D Electrophoresis Modeling Of Multienzyme Cutting Of Polypeptides, Howard Mayes

Theses

2-Dimensional Electrophoresis is one of the tools in the identification of proteins by molecular weight and pH. The display of molecular weight allows the researcher to quickly identify whether a specific protein or peptide string is in the sample. The pH measurement allows even better resolution between different species in the sample. The MultiEnzyme ElectroPhoresis (MEEP) program tries to model that by providing a graph that displays separated protein strings by both molecular weight and pH. The ability to cleave the protein with 43 different enzyme variations allows the researcher to analyze appropriate enzymes to isolate a protein subsequence before ...


Tension Directly Stabilizes Reconstituted Kinetochore-Microtubule Attachments, Bungo Akiyoshi, Krishna K. Sarangapani, Andrew F. Powers, Christian R. Nelson, Steve Reichow, Hugo Arellano-Santoyo, Tamir Gonen, Jeffrey A. Ranish, Charles L. Asbury, Sue Biggins Oct 2017

Tension Directly Stabilizes Reconstituted Kinetochore-Microtubule Attachments, Bungo Akiyoshi, Krishna K. Sarangapani, Andrew F. Powers, Christian R. Nelson, Steve Reichow, Hugo Arellano-Santoyo, Tamir Gonen, Jeffrey A. Ranish, Charles L. Asbury, Sue Biggins

Steve Reichow

Kinetochores are macromolecular machines that couple chromosomes to dynamic microtubule tips during cell division, thereby generating force to segregate the chromosomes. Accurate segregation depends on selective stabilization of correct ‘bi-oriented’ kinetochore-microtubule attachments, which come under tension due to opposing forces exerted by microtubules. Tension is thought to stabilize these bi-oriented attachments indirectly, by suppressing the destabilizing activity of a kinase, Aurora B. However, a complete mechanistic understanding of the role of tension requires reconstitution of kinetochore-microtubule attachments for biochemical and biophysical analyses in vitro. Here we show that native kinetochore particles retaining the majority of kinetochore proteins can be purified ...


Detecting And Accounting For Multiple Sources Of Positional Variance In Peak List Registration Analysis And Spin System Grouping, Andrey Smelter, Eric C. Rouchka, Hunter N. B. Moseley Aug 2017

Detecting And Accounting For Multiple Sources Of Positional Variance In Peak List Registration Analysis And Spin System Grouping, Andrey Smelter, Eric C. Rouchka, Hunter N. B. Moseley

Molecular and Cellular Biochemistry Faculty Publications

Peak lists derived from nuclear magnetic resonance (NMR) spectra are commonly used as input data for a variety of computer assisted and automated analyses. These include automated protein resonance assignment and protein structure calculation software tools. Prior to these analyses, peak lists must be aligned to each other and sets of related peaks must be grouped based on common chemical shift dimensions. Even when programs can perform peak grouping, they require the user to provide uniform match tolerances or use default values. However, peak grouping is further complicated by multiple sources of variance in peak position limiting the effectiveness of ...


Real-Time Sensing Of Single-Ligand Delivery With Nanoaperture-Integrated Microfluidic Devices, W. Elliott Martin, Ning Ge, Bernadeta R. Srijanto, Emily Furnish, C. Patrick Collier, Christine A. Trinkle, Christopher I. Richards Jul 2017

Real-Time Sensing Of Single-Ligand Delivery With Nanoaperture-Integrated Microfluidic Devices, W. Elliott Martin, Ning Ge, Bernadeta R. Srijanto, Emily Furnish, C. Patrick Collier, Christine A. Trinkle, Christopher I. Richards

Chemistry Faculty Publications

The measurement of biological events on the surface of live cells at the single-molecule level is complicated by several factors including high protein densities that are incompatible with single-molecule imaging, cellular autofluorescence, and protein mobility on the cell surface. Here, we fabricated a device composed of an array of nanoscale apertures coupled with a microfluidic delivery system to quantify single-ligand interactions with proteins on the cell surface. We cultured live cells directly on the device and isolated individual epidermal growth factor receptors (EGFRs) in the apertures while delivering fluorescently labeled epidermal growth factor. We observed single ligands binding to EGFRs ...


Use Of Crispr To Develop A Follicle Stimulating Hormone Receptor Knockout In Human Granulosa Cells To Study Lipid Raft Residency, Jenna Pradhuman Jun 2017

Use Of Crispr To Develop A Follicle Stimulating Hormone Receptor Knockout In Human Granulosa Cells To Study Lipid Raft Residency, Jenna Pradhuman

Honors Theses

Human follicle stimulating hormone (hFSH) is a protein hormone responsible for stimulating the gonads and is necessary for regulating reproductive systems in both females and males. The actions of hFSh are carried out by the hFSH receptor (hFSHR), a seven transmembrane receptor that belongs to the G protein-coupled receptor family. Once FSH activates its receptor, the G protein inside the cell that is associated with the receptor is activated and starts a cascade of signaling that results in activation of protein kinase A (PKA) and the p44/42 MAP kinase (MAPK). The activation of these secondary proteins is responsible for ...


Augmenting The Anisotropic Network Model With Torsional Potentials Improves Path Performance, Enabling Detailed Comparison With Experimental Rate Data, Srinivas Niranj Chandrasekaran, Charles W. Carter Jr Feb 2017

Augmenting The Anisotropic Network Model With Torsional Potentials Improves Path Performance, Enabling Detailed Comparison With Experimental Rate Data, Srinivas Niranj Chandrasekaran, Charles W. Carter Jr

Open Access Articles

PATH algorithms for identifying conformational transition states provide computational parameters-time to the transition state, conformational free energy differences, and transition state activation energies-for comparison to experimental data and can be carried out sufficiently rapidly to use in the "high throughput" mode. These advantages are especially useful for interpreting results from combinatorial mutagenesis experiments. This report updates the previously published algorithm with enhancements that improve correlations between PATH convergence parameters derived from virtual variant structures generated by RosettaBackrub and previously published kinetic data for a complete, four-way combinatorial mutagenesis of a conformational switch in Tryptophanyl-tRNA synthetase.


Improving The Usage Of Unnatural Amino Acids In Proteins: Thioamides And Other Biophysical Probes, Christopher Russell Walters Jan 2017

Improving The Usage Of Unnatural Amino Acids In Proteins: Thioamides And Other Biophysical Probes, Christopher Russell Walters

Publicly Accessible Penn Dissertations

Methods for genetically and synthetically manipulating protein composition enable a greater flexibility in the study of protein dynamics and function. However, current techniques for incorporating biophysical probes in the form of unnatural amino acids (Uaas) can suffer from poor yield, limited selectivity for the desired probe, and an insufficient understanding of the impact that the probe has on protein structure and function. Each of the studies discussed herein addresses one or more of these shortcomings in an effort to improve the usage of Uaas in biochemistry. We have shown that using inteins as traceless, cleavable purification tags enables the separation ...


Purification, Optimization, And Growth Of New Delhi Metallo-Β-Lactamase-1 Protein Crystals Mixed With Nz218 Inhibitor, Brandon M. Wills May 2016

Purification, Optimization, And Growth Of New Delhi Metallo-Β-Lactamase-1 Protein Crystals Mixed With Nz218 Inhibitor, Brandon M. Wills

Celebration of Learning

New Delhi metallo-β-lactamase-1 is a problematic gene found in certain strains of bacteria that cause them to become antibiotic resistant to nearly all known antibiotics. While some antibiotics are available to treat patients with a bacterial infection, most are toxic or do not have 100% success rates. With that being said, it is imperative that we search for a molecule that is successfully able to inhibit the effects of this gene every time. Such a discovery would help tremendously with new antibiotic drug development and also prevent further damage by these dangerous bacteria. In this presentation, I will describe the ...


Using Simple Self-Assembling Peptides To Attain Novel Protein-Like Functions, Tyler Smith May 2016

Using Simple Self-Assembling Peptides To Attain Novel Protein-Like Functions, Tyler Smith

Syracuse University Honors Program Capstone Projects

Proteins carry out many extremely efficient functions, including catalysis and biomolecule recognition. Underlying this efficiency is their extraordinary complexity and ability to fold into unique three-dimensional structures. Attempts to replicate this efficiency through de novo design have only shown moderate success, and it is unclear how modern-day proteins may have evolved. However, short peptides that alternate hydrophobic and hydrophilic residues can self-assemble into amyloid fibrils to achieve well-defined secondary structure. These aggregates may have served as a template from which the first proteins were derived. We designed self-assembling seven-residue peptides that are able to act as Zn2+-dependent esterases ...


Engineered Protein Polymer-Gold Nanoparticle Hybrid Materials For Small Molecule Delivery, Min Dai, Ja Frezzo, E Sharma, R Chen, N Singh, C Yuvienco, E Caglar, S Xiao, A Saxena, Jk Montclare Feb 2016

Engineered Protein Polymer-Gold Nanoparticle Hybrid Materials For Small Molecule Delivery, Min Dai, Ja Frezzo, E Sharma, R Chen, N Singh, C Yuvienco, E Caglar, S Xiao, A Saxena, Jk Montclare

Publications and Research

We have fabricated protein polymer-gold nanoparticle (P-GNP) nanocomposites that exhibit enhanced binding and delivery properties of the small hydrophobic molecule drug, curcumin, to the model breast cancer cell line, MCF-7. These hybrid biomaterials are constructed via in situ GNP templated-synthesis with genetically engineered histidine tags. The P-GNP nanocomposites exhibit enhanced small molecule loading, sustained release and increased uptake by MCF-7 cells. When compared to the proteins polymers alone, the P-GNPs demonstrate a greater than 7-fold increase in curcumin binding, a nearly 50% slower release profile and more than 2-fold increase in cellular uptake of curcumin. These results suggest that P-GNP ...


The Application Of Hydrogen/Deuterium Exchange And Covalent Labeling Coupled With Mass Spectrometry To Examine Protein Structure, Nicholas B. Borotto Jan 2016

The Application Of Hydrogen/Deuterium Exchange And Covalent Labeling Coupled With Mass Spectrometry To Examine Protein Structure, Nicholas B. Borotto

Doctoral Dissertations

Thorough insight into a protein’s structure is necessary to understand how it functions and what goes wrong when it malfunctions. The structure of proteins, however, is not easily analyzed. The analysis must take place under a narrow range of conditions or risk perturbing the very structure being probed. Furthermore, the wide diversity in size and chemistry possible in proteins significantly complicates this analysis. Despite this numerous methods have been developed in order to analyze protein structure. In this work, we demonstrate that mass spectrometry (MS)-based techniques are capable of characterizing the structure of particularly challenging proteins. This is ...


Characterization Of A Protein: Protein Interaction Between Rnf181 And The Platelet Integrin, Αiibβ3, Seamus Allen Nov 2015

Characterization Of A Protein: Protein Interaction Between Rnf181 And The Platelet Integrin, Αiibβ3, Seamus Allen

PhD theses

The integrin αIIbβ3 is a heterodimeric protein that exists in an inactive, resting state within the membrane of quiescent platelets. In response to platelet activation within a pro-thrombotic environment, the integrin αIIbβ3 becomes active, serves as a receptor for a number of ligands including fibrinogen, and orientates platelet – platelet cohesion. The αIIb and β3 subunits maintain the integrin in a resting conformational state by forming a tight clasp at their cytoplasmic tails (CT’s). Disruption of this clasp by platelet activation initiates the transition of the integrin from an inactive conformation to an active ligand binding state. The highly conserved ...


Cell Type–Dependent Mechanisms For Formin-Mediated Assembly Of Filopodia, Lorna E. Young, Ernest G. Heimsath, Henry N. Higgs Oct 2015

Cell Type–Dependent Mechanisms For Formin-Mediated Assembly Of Filopodia, Lorna E. Young, Ernest G. Heimsath, Henry N. Higgs

Open Dartmouth: Faculty Open Access Scholarship

Filopodia are finger-like protrusions from the plasma membrane and are of fundamental importance to cellular physiology, but the mechanisms governing their assembly are still in question. One model, called convergent elongation, proposes that filopodia arise from Arp2/3 complex-nucleated dendritic actin networks, with factors such as formins elongating these filaments into filopodia. We test this model using constitutively active constructs of two formins, FMNL3 and mDia2. Surprisingly, filopodial assembly requirements differ between suspension and adherent cells. In suspension cells, Arp2/3 complex is required for filopodial assembly through either formin. In contrast, a subset of filopodia remains after Arp2/3 ...


Binding Interactions Of (R)- And (S)-Hydroxypropyl-Com Dehydrogenases And The Zinc Knuckle Proteins Air1 And Air2, Jeremy W. Bakelar May 2015

Binding Interactions Of (R)- And (S)-Hydroxypropyl-Com Dehydrogenases And The Zinc Knuckle Proteins Air1 And Air2, Jeremy W. Bakelar

All Graduate Theses and Dissertations

This work is focused on understanding protein function by describing how paralogous proteins with overlapping and distinct functions interact with their substrates and with other proteins. Two model systems are the subject of this research: (1) the stereospecific dehydrogenases R- and S-HPCDH, and (2) the zinc knuckle proteins Air1 and Air2.

R- and S-HPCDH are homologous enzymes that are central to the metabolism of propylene and epoxide in the soil bacterium Xanthobacter autotrophicus. The bacterium produces R- and S-HPCDH simultaneously to facilitate transformation of R- and S-enantiomers of epoxypropane to a common achiral product 2-ketopropyl-CoM (2-KPC ...


Distinguishing Macrophage Activation States By Mass Spectrometry, Matthias Manfred Knust May 2015

Distinguishing Macrophage Activation States By Mass Spectrometry, Matthias Manfred Knust

Theses and Dissertations

Macrophages are versatile and highly adaptive cells that are involved in a wide range of physiological processes including host defense, homeostasis or regeneration, as well as pathogenesis. They react to their microenvironment, assuming various roles based on chemical and/or physical cues, and can reversibly shift between these so-called activation states. Concurrently, the technique of immunohistochemistry is used to gain spatial information on activated macrophages on tissue sections. The aim of this work was to find mass spectral biomarkers that allow the differentiation of activation states, and establish conditions that can be used in imaging mass spectrometry (IMS) experiments to ...


Nonenzymatic Glycosylation Of Erythrocyte Membrane Proteins. Relevance To Diabetes, J A. Miller, Ellen M. Gravallese, H F. Bunn Apr 2015

Nonenzymatic Glycosylation Of Erythrocyte Membrane Proteins. Relevance To Diabetes, J A. Miller, Ellen M. Gravallese, H F. Bunn

Ellen M. Gravallese

Nonenzymatic glycosylation of proteins of the erythrocyte membrane was determined by incubating erythrocyte ghosts with [3H]borohydride. The incorporation of tritium into protein provides a reliable assay of ketoamine linkages. The membrane proteins from 18 patients with diabetes incorporated twice as much radioactivity as membrane proteins from normal erythrocytes. After acid hydrolysis, amino acid analysis showed that the majority of radioactivity was localized to glucosyllysine. Autoradiograms showed that all of the major proteins of the erythrocyte membrane, separated by electrophoresis on sodium dodecyl sulfate gels, contained ketoamine linkages. No protein bands in either normal or diabetic erythrocytes showed significant preferential ...


Morphological Transformations In The Magnetite Biomineralizing Protein Mms6 In Iron Solutions: A Small-Angle X‑Ray Scattering Study, Honghu Zhang, Xunpei Liu, Shuren Feng, Wenjie Wang, Klaus Schmidt-Rohr, Mufit Akinc, Marit Nilsen-Hamilton, David Vaknin, Surya K. Mallapragada Feb 2015

Morphological Transformations In The Magnetite Biomineralizing Protein Mms6 In Iron Solutions: A Small-Angle X‑Ray Scattering Study, Honghu Zhang, Xunpei Liu, Shuren Feng, Wenjie Wang, Klaus Schmidt-Rohr, Mufit Akinc, Marit Nilsen-Hamilton, David Vaknin, Surya K. Mallapragada

Chemical and Biological Engineering Publications

Magnetotactic bacteria that produce magnetic nanocrystals of uniform size and well-defined morphologies have inspired the use of biomineralization protein Mms6 to promote formation of uniform magnetic nanocrystals in vitro. Small angle X-ray scattering (SAXS) studies in physiological solutions reveal that Mms6 forms compact globular threedimensional (3D) micelles (approximately 10 nm in diameter) that are, to a large extent, independent of concentration. In the presence of iron ions in the solutions, the general micellar morphology is preserved, however, with associations among micelles that are induced by iron ions. Compared with Mms6, the m2Mms6 mutant (with the sequence of hydroxyl/carboxyl containing ...


Liver Perilipin 5 Expression Worsens Hepatosteatosis But Not Insulin Resistance In High Fat-Fed Mice, Michelle B. Trevino, David Mazur-Hart, Yui Machida, Timothy King, Joseph Nadler, Elena V. Galkina, Arjun Poddar, Sucharita Dutta, Yumi Imai Jan 2015

Liver Perilipin 5 Expression Worsens Hepatosteatosis But Not Insulin Resistance In High Fat-Fed Mice, Michelle B. Trevino, David Mazur-Hart, Yui Machida, Timothy King, Joseph Nadler, Elena V. Galkina, Arjun Poddar, Sucharita Dutta, Yumi Imai

Mathematics & Statistics Faculty Publications

Perilipin 5 (PLIN5) is a lipid droplet (LD) protein highly expressed in oxidative tissues, including the fasted liver. However, its expression also increases in nonalcoholic fatty liver. To determine whether PLIN5 regulates metabolic phenotypes of hepatosteatosis under nutritional excess, liver targeted overexpression of PLIN5 was achieved using adenoviral vector (Ad-PLIN5) in male C57BL/6J mice fed high-fat diet. Mice treated with adenovirus expressing green fluorescent protein (GFP) (Ad-GFP) served as control. Ad-PLIN5 livers increased LD in the liver section, and liquid chromatography with tandem mass spectrometry revealed increases in lipid classes associated with LD, including triacylglycerol, cholesterol ester, and phospholipid ...


The Effect Of O Antigen Loss On The Protein Composition And Inflammatory Response Elicited By Klebsiella Pneumoniae, Bethaney Cahill Jan 2015

The Effect Of O Antigen Loss On The Protein Composition And Inflammatory Response Elicited By Klebsiella Pneumoniae, Bethaney Cahill

UNF Graduate Theses and Dissertations

Klebsiella pneumoniae is a Gram-negative pathogen associated with numerous infections. Like all Gram-negative bacteria, K. pneumoniae naturally release outer membrane vesicles (OMVs) during all stages of cellular growth. OMVs are composed of the outer membrane components such as lipopolysaccharide (LPS) and outer membrane proteins and contain cytosolic and periplasmic proteins in the lumen. K. pneumoniae is often found to lack an O antigen. The absence of the O antigen has been reported to alter the protein content of the membrane which may further alter the immune response elicited by K. pneumoniae. Therefore the purpose of this study was to analyze ...


The Formin Fmnl3 Assembles Plasma Membrane Protrusions That Participate In Cell–Cell Adhesion, Timothy J. Gauvin, Lorna E. Young, Henry N. Higgs Nov 2014

The Formin Fmnl3 Assembles Plasma Membrane Protrusions That Participate In Cell–Cell Adhesion, Timothy J. Gauvin, Lorna E. Young, Henry N. Higgs

Open Dartmouth: Faculty Open Access Scholarship

FMNL3 is a vertebrate-specific formin protein previously shown to play a role in angiogenesis and cell migration. Here we define the cellular localization of endogenous FMNL3, the dynamics of GFP-tagged FMNL3 during cell migration, and the effects of FMNL3 suppression in mammalian culture cells. The majority of FMNL3 localizes in a punctate pattern, with >95% of these puncta being indistinguishable from the plasma membrane by fluorescence microscopy. A small number of dynamic cytoplasmic FMNL3 patches also exist, which enrich near cell–cell contact sites and fuse with the plasma membrane at these sites. These cytoplasmic puncta appear to be part ...


Automatic Animation Of Molecular Motion Using Python And Cinema 4d, Diana Zajac, Nathaniel Smith, Dan Gurnon Nov 2014

Automatic Animation Of Molecular Motion Using Python And Cinema 4d, Diana Zajac, Nathaniel Smith, Dan Gurnon

Science Research Fellows Posters

No abstract provided.


Early Folding Biases In The Folding Free-Energy Surface Of Βα-Repeat Proteins: A Dissertation, Robert P. Nobrega Jul 2014

Early Folding Biases In The Folding Free-Energy Surface Of Βα-Repeat Proteins: A Dissertation, Robert P. Nobrega

GSBS Dissertations and Theses

Early events in folding can determine if a protein is going to fold, misfold, or aggregate. Understanding these deterministic events is paramount for de novo protein engineering, the enhancement of biopharmaceutical stabilities, and understanding neurodegenerative diseases including amyotrophic lateral sclerosis and Alzheimer's disease. However, the physicochemical and structural biases within high energy states of protein biopolymers are poorly understood.

A combined experimental and computational study was conducted on the small β/α-repeat protein CheY to determine the structural basis of its submillisecond misfolding reaction to an off-pathway intermediate. Using permutations, we were able to discriminate between the roles of ...


Expression In Yeast Of The Metacaspase Scp3 From The Fungus Schizophyllum Commune, Anne Kaminski Jun 2014

Expression In Yeast Of The Metacaspase Scp3 From The Fungus Schizophyllum Commune, Anne Kaminski

Honors Theses

Metacaspases are caspase homologs that are found in plants, fungi, and protists. They are involved in programmed cell death, a structured and regulated process to break down cells. There is limited information available on metacaspases and further research is necessary to understand how they work. One reason metacaspases are studied is that they are considered to be a potential drug target for pathogenic microorganisms. Five metacaspases scp1-5 were identified in S. commune through their similarity to the yeast metacaspase Yca1. The overall goal of the lab is to study and characterize these proteins. This thesis outlines the path taken to ...


Stoichiometries And Affinities Of Interacting Proteins From Concentration Series Of Solution Scattering Data: Decomposition By Least Squares And Quadratic Optimization, Himanshu Chandola, Tim E. Williamson, Bruce A. Craig, Alan M. Friedman, Chris Bailey-Kellogg Mar 2014

Stoichiometries And Affinities Of Interacting Proteins From Concentration Series Of Solution Scattering Data: Decomposition By Least Squares And Quadratic Optimization, Himanshu Chandola, Tim E. Williamson, Bruce A. Craig, Alan M. Friedman, Chris Bailey-Kellogg

Open Dartmouth: Faculty Open Access Scholarship

In studying interacting proteins, complementary insights are provided by analyzing both the association model (the stoichiometry and affinity constants of the intermediate and final complexes) and the quaternary structure of the resulting complexes. Many current methods for analyzing protein interactions either give a binary answer to the question of association and no information about quaternary structure or at best provide only part of the complete picture. Presented here is a method to extract both types of information from X-ray or neutron scattering data for a series of equilibrium mixtures containing the initial components at different concentrations. The method determines the ...


A Novel Selection Technology For The Discovery Of High-Affinity Human Proteins, David Busha Jan 2014

A Novel Selection Technology For The Discovery Of High-Affinity Human Proteins, David Busha

Molecular, Cellular, and Developmental Biology Graduate Theses & Dissertations

Proteins that bind with high-affinity to cellular targets can be useful therapeutic treatments. High-throughput affinity screening of large protein libraries is often more successful at discovering novel high-affinity proteins than rational-design approaches. Display techniques such and phage and yeast display are commonly used in this screening process. However, bacteria and yeast cells can misfold or otherwise inappropriately express mammalian and human proteins due to differences in codon usage, protein folding machinery and post-translational modifications. Therefore, a display system that is entirely based in human cells could aid in the discovery of novel, high-affinity proteins. No such system has been described ...