Biochemistry, Biophysics, and Structural Biology Commons^™

Computational And Experimental Investigation Into The Determinants Of Protein Structure, Folding, And Stability In The Β-Grasp Superfamily, John T. Bedford Ii

Chemistry & Biochemistry Theses & Dissertations

Elucidating the mechanisms of protein folding and unfolding is one of the greatest scientific challenges in basic science. The overarching goal is to predict three-dimensional structures from their amino acid sequences. Understanding the determinants of protein folding and stability can be facilitated through the study of evolutionarily related but diverse proteins. Insights can also be gained through the study of proteins from extremophiles that may more closely resemble the primordial proteins. In this doctoral research, three aims were accomplished to characterize the structure, folding and unfolding behavior within the β-grasp superfamily. We propose that the determinants of structure, stability, and …

Toward The Discovery Of Biological Functions Associated With The Mechanosensor Mtl1p Of Saccharomyces Cerevisiae Via Integrative Multi-Omics Analysis, Nelson Martínez-Matías, Nataliya Chorna, Sahily González-Crespo, Lilliam Villanueva, Ingrid Montes-Rodríguez, Loyda M. Melendez-Aponte, Abiel Roche-Lima, Kelvin Carrasquillo-Carrión, Ednalise Santiago-Cartagena, Brian C. Rymond, Mohan Babu, Igor Stagljar, José R. Rodríguez-Medina

Biology Faculty Publications

Functional analysis of the Mtl1 protein in Saccharomyces cerevisiae has revealed that this transmembrane sensor endows yeast cells with resistance to oxidative stress through a signaling mechanism called the cell wall integrity pathway (CWI). We observed upregulation of multiple heat shock proteins (HSPs), proteins associated with the formation of stress granules, and the phosphatase subunit of trehalose 6-phosphate synthase which suggests that mtl1Δ strains undergo intrinsic activation of a non-lethal heat stress response. Furthermore, quantitative global proteomic analysis conducted on TMT-labeled proteins combined with metabolome analysis revealed that mtl1Δ strains exhibit decreased levels of metabolites of carboxylic acid metabolism, decreased …

Construction Of E. Coli Strain With N-Terminal Domain Deletion In The Biotin Carboxyl Carrier Protein, Dara Khosravi

Honors Capstones

No abstract provided.

Protocol For Rapid Assessment Of The Efficacy Of Novel Wnt Inhibitors Using Zebrafish Models, Meghan G. Haney, Mary Wimsett, Chunming Liu, Jessica S. Blackburn

Molecular and Cellular Biochemistry Faculty Publications

Dysregulation of Wnt signaling is a hallmark of many cancers, and the development of effective, non-toxic small-molecule Wnt inhibitors is desirable. Off-target toxicities of new compounds are typically tested in mouse models, which is both costly and time consuming. Here, we present a rapid and inexpensive protocol to determine the in vivo toxicity and efficacy of novel Wnt inhibitors in zebrafish using a combination of a fluorescence reporter assay as well as eye rescue and fin regeneration assays. These experiments are completed within 1 week to rapidly narrow drug candidates before moving to more expensive pre-clinical testing.

For complete details …

Biophysical Characterization Of The Par-4 Tumor Suppressor: Evidence Of Structure Outside The Coiled Coil Domain And Interactions With Platinum Chemotherapeutics, Andrea Megan Clark

Chemistry & Biochemistry Theses & Dissertations

Prostate apoptosis response-4 (Par-4) is an apoptosis-inducing tumor suppressor protein. Full-length Par-4 has previously been shown to be a predominantly intrinsically disordered protein (IDP) under neutral conditions, with significant regular secondary structure evident only within the C-terminal coiled coil domain. However, IDPs can gain ordered structure through the process of induced folding, which often occurs under non-neutral conditions. Previous work has shown that the Par-4 leucine zipper, which is a subset of the C-terminal coiled coil domain, is disordered under neutral conditions, but forms a dimeric coiled coil at acidic pH. Increase in ionic strength was also shown to increase …

Single-Cell Transcriptomics Identifies Limbal Stem Cell Population And Cell Types Mapping Its Differentiation Trajectory In Limbal Basal Epithelium Of Human Cornea, De-Quan Li, Sangbae Kim, Jin-Miao Li, Qianmiao Gao, Jongsu Choi, Fang Bian, Jiaoyue Hu, Yun Zhang, Jin Li, Rong Lu, Yumei Li, Stephen C Pflugfelder, Hongyu Miao, Rui Chen

Faculty, Staff and Students Publications

PURPOSE: This study aimed to uncover novel cell types in heterogenous basal limbus of human cornea for identifying LSC at single cell resolution.

METHODS: Single cells of human limbal basal epithelium were isolated from young donor corneas. Single-cell RNA-Sequencing was performed using 10x Genomics platform, followed by clustering cell types through the graph-based visualization method UMAP and unbiased computational informatic analysis. Tissue RNA in situ hybridization with RNAscope, immunofluorescent staining and multiple functional assays were performed using human corneas and limbal epithelial culture models.

RESULTS: Single-cell transcriptomics of 16,360 limbal basal cells revealed 12 cell clusters belonging to three lineages. …

Mitochondrial Transfer From Mesenchymal Stem Cells Improves Neuronal Metabolism After Oxidant Injury In Vitro: The Role Of Miro1, Nancy Tseng, Scott C Lambie, Christopher Q Huynh, Bridget Sanford, Manisha Patel, Paco S Herson, D Ryan Ormond

Faculty, Staff and Students Publications

Stroke-induced cerebral ischemia is a major cause of death and disability. The disruption of blood flow results in neuronal and glial cell death leading to brain injury. Reperfusion restores oxygen to the affected tissue, but can also cause damage through an enhanced oxidative stress and inflammatory response. This study examines mitochondrial transfer from MSC to neurons and the role it plays in neuronal preservation after oxidant injury. We observed the transfer of mitochondria from MSC to mouse neurons in vitro following hydrogen peroxide exposure. The observed transfer was dependent on cell-to-cell contact and led to increased neuronal survival and improved …

The Impact Of Covid-19 On Kidney Transplantation And The Kidney Transplant Recipient – One Year Into The Pandemic, Pascale Khairallah, Nidhi Aggarwal, Ahmed A Awan, Chandan Vangala, Medha Airy, Jenny S Pan, Bhamidipati V R Murthy, Wolfgang C Winkelmayer, Venkat Ramanathan

Faculty, Staff and Students Publications

The COVID‐19 pandemic has significantly changed the landscape of kidney transplantation in the United States and worldwide. In addition to adversely impacting allograft and patient survival in postkidney transplant recipients, the current pandemic has affected all aspects of transplant care, including transplant referrals and listing, organ donation rates, organ procurement and shipping, and waitlist mortality. Critical decisions were made during this period by transplant centers and individual transplant physicians taking into consideration patient safety and resource utilization. As countries have begun administering the COVID vaccines, new and important considerations pertinent to our transplant population have arisen. This comprehensive review focuses …

Lions, Tigers, And Hemes - Oh My! A Dynamic Look At The Electronic Effects Of Porphyrin Substitution On Cytochrome P450 Olet, Alexis J. Holwerda

Senior Theses

OleT, a member of the CYP152 family of cytochrome P450s (CYPs), decarboxylates fatty acids using hydrogen peroxide as an oxidant. The resultant products are a terminal alkene and carbon dioxide. This C–C cleavage reaction is highly atypical for CYPs, which prototypically oxygenate substrates, and provides a potential means to enzymatically produce drop-in fuels. OleT contains a heme-iron cofactor that facilitates decarboxylation through the activation of hydrogen peroxide. The catalytic cycle, as determined by transient kinetics, includes two ferryl intermediates known as Compound I (Ole-I) and Compound II (Ole-II). Ole-I performs substrate hydrogen abstraction and subsequent single electron transfer to Ole-II …

The Effect Of A Folate Free Diet On Tumor Burden And Chemotherapy Effectiveness, Kendall E. Stalls

Senior Theses

Our research is to understand the effect of the microbiota on colorectal cancer. Evidence has been shown that the microbiota is linked to the development and progression of colorectal cancer. The research focuses on how the gut microbiome influences the effectiveness of colorectal cancer treatments. Specifically, how the composition of the microbiota and the metabolites that it secretes changes the response to chemotherapy. We also research how tumor burden can be altered through changing the composition of the gut bacteria. Our experiments are performed on APC/Min+ mice, that have been genetically altered to have colon cancer. We conducted an experiment …

Purification And Functional Characterization Of The Iron-Responsive Transcription Factor Aft1 From C. Glabrata, Jade Ikahihifo-Bender

Senior Theses

Due to its unique ability to serve as both an electron donor and acceptor, iron is utilized as a co-factor for many biological processes, including electron transfer, oxygen binding, and vitamin synthesis. Iron is also a key factor during fungal infections as the human host and invading pathogens battle over limited iron pools. The primary iron-responsive transcription factor Aft1 in the opportunistic pathogenic yeast Candida glabrata responds to iron deficiency by activating expression of iron acquisition genes. However, the mechanisms for sensing intracellular iron levels and regulating Aft1 activity in response to iron are unknown. The C. glabrata iron regulation …

A Spatially Explicit Model Of Stabilizing Selection For Improving Phylogenetic Inference, Jeremy M. Beaulieu, Brian C. O'Meara, Michael A. Gilchrist

Biological Sciences Faculty Publications and Presentations

Ultraconserved elements (UCEs) are stretches of hundreds of nucleotides with highly conserved cores flanked by variable regions. Although the selective forces responsible for the preservation of UCEs are unknown, they are nonetheless believed to contain phylogenetically meaningful information from deep to shallow divergence events. Phylogenetic applications of UCEs assume the same degree of rate heterogeneity applies across the entire locus, including variable flanking regions. We present a Wright-Fisher model of selection on nucleotides (SeION) which includes the effects of mutation, drift, and spatially varying, stabilizing selection for an optimal nucleotide sequence. The SeION model assumes the strength of stabilizing selection …

Evaluating The Role Of The Stringent Response Mechanism In Clostridioides Difficile Survival And Pathogenesis, Astha Pokhrel

Chemistry & Biochemistry Theses & Dissertations

The human pathogen Clostridioides difficile is increasingly tolerant of multiple antibiotics and causes infections with a high rate of recurrence, creating an urgent need for new preventive and therapeutic strategies. The stringent response, a universal bacterial response to extracellular stresses, governs antibiotic survival and pathogenesis in diverse organisms but has not previously been characterized in C. difficile. This dissertation explores the ability of C. difficile to mount the stringent response. The bacteria encode a full-length, canonical bifunctional Rel/Spo Homolog or RSH enzyme. C. difficile RSH is incapable of utilizing GTP as a substrate but readily synthesizes putative 5’-pGpp-3’ alarmones. …

Modeling Of Aqueous Root Extract Compounds Of Ruellia Tuberosa L. For Alpha-Glucosidase Inhibition Through In Silico Study, Anna Safitri, Dewi Ratih Tirto Sari, Fatchiyah Fatchiyah, Anna Roosdiana

Makara Journal of Science

This study aims to analyze the inhibitory activities of aqueous root extract compounds of Ruellia tuberosa L. toward alpha-glucosidase protein by computational docking analysis. Three major compounds contained in the extracts (i.e., betaine, daidzein, and hispidulin) were selected as ligands; quercetin and acarbose were used as the reference. Computational docking analysis was performed using the HEX 8.0.0 program and visualized using the Discovery Studio Visualizer v19.1.0.18287 (2019 version) on the basis of the scoring functions. The interactions between ligands and alpha-glucosidase protein showed different binding patterns. The types of bonds involved in the interaction between the enzyme and these ligands …

Morph- And Sex-Specific Differences In Corticosterone Of The Arizona Tiger Salamander (Ambystoma Mavortium Nebulosum), Megan Zerger

Scholars Week

Life history morph, sex, and body condition are traits that may influence stress within salamander populations because of differences in physiology and environmental conditions. Given widespread declines and the effects chronic stress can have on amphibian health, it is important to understand within-population drivers of stress and how population level variation may influence population viability. Thus, the objective of our study was to assess how corticosterone varies within the Arizona tiger salamander (Ambystoma mavortium nebulosum) population at the Mexican Cut Nature Preserve. We used a non-invasive skin swabbing method to collect baseline and elevated corticosterone from paedomorph (aquatic …

Biological Activity And Solubility Of 5-Methoxy-1,4-Benzoquinone Having Bromoheptyl And Bromodecyl Substituents In The N-Octanol/Water System, Siti Mariyah Ulfa, Fath Dwisari, Laras Pangesti, Mohammad Farid Rahman

Makara Journal of Science

The biological activity and solubility of compounds are influenced by its chemical structure. These properties can be improved by substituting alkyl, alkoxy, and/or haloalkane in the parent skeleton. In this research, the synthesis of 3-(7-bromoheptyl)-2-methyl-5-methoxy-1,4-benzoquinone (3a) and 3-(10-bromodecyl)-2-methyl-5-methoxy-1,4-benzoquinone (3b) was achieved through the decarboxylation reaction. The solubility and biological activity of 3a and 3b were compared with that of thymoquinone (TQ), which acts as an anti-inflammatory agent. Compounds 3a and 3b were successfully synthesized and analyzed using Fourier Transform Infra-Red (FTIR) and Nuclear Magnetic Resonance (NMR). The FTIR spectrum showed the increasing intensity of …

Case 32: Inhibition Of Protein Tyrosine Phosphatase Enzymes As A Possible Treatment For Diabetes, Dr. Kathleen Cornely

Chemistry & Biochemistry Case Studies

Protein tyrosine phosphatases (PTPs) are investigated because of their ability to reverse the effects of protein tyrosine kinases (PTKs). Inhibitors of the PTP investigated in this study have insulin-mimetic effects and have potential in the treatment of diabetes.

Analysis Of Heavy Metal In Hair Samples Using Icp Oes, Emma Suvacarov

Honors Scholars & Undergraduate Research Poster Symposium Programs

Exposure to heavy metals such as mercury, lead, and arsenic can negatively affect the structure and function of cells, tissues, and organs. Such heavy metals can be excreted from the hair as metabolic products during its growth process thus hair analysis is thought to be a reliable indicator for the environmental accumulation of heavy metals. Our research aims to develop a methodology to quantify heavy metal concentrations found in biological samples using Inductively Coupled Plasma Spectroscopy. Current research involves preparing hair samples for analysis and creating calibration curves for heavy metals through a process that is accurate, precise, and easily …

Lighting The Way: Recent Insights Into The Structure And Regulation Of Phototropin Blue Light Receptors, Jaynee E. Hart, Kevin H. Gardner

Publications and Research

The phototropins (phots) are light-activated kinases that are critical for plant physiology and the many diverse optogenetic tools that they have inspired. Phototropins combine two bluelight- sensing Light–Oxygen–Voltage (LOV) domains (LOV1 and LOV2) and a C-terminal serine/threonine kinase domain, using the LOV domains to control the catalytic activity of the kinase. While much is known about the structure and photochemistry of the light-perceiving LOV domains, particularly in how activation of the LOV2 domain triggers the unfolding of alpha helices that communicate the light signal to the kinase domain, many questions about phot structure and mechanism remain. Recent studies have made …

Insights Into Viral Genome Function Through Comparative Structural Analysis, Lydia Phillips

UNO Student Research and Creative Activity Fair

Enteroviruses are single stranded RNA viruses which have caused many public health concerns, particularly in children. These viruses are responsible for polio, hand, foot, and mouth disease, many polio-like neurological diseases, and the common cold. The enterovirus called Coxsackievirus B3 (CVB3), a close relative to poliovirus, has been shown responsible for severe human disease, including pancreatitis, myocarditis, and type 1 diabetes. A unique advantage of studying CVB3 is the existence of a naturally occurring strain (CVB3/GA) which displays no known pathogenicity. Earlier comparative genomic studies have shown that the primary difference between avirulent CVB3/GA and the virulent strains of CVB3 …

Latitude Dictates Plant Diversity Effects On Instream Decomposition, Luz Boyero, Javier Pérez, Naiara López-Rojo, Alan M. Tonin, Francisco Correa-Araneda, Richard G. Pearson, Jaime Bosch, Ricardo J. Albariño, Sankarappan Anbalagan, Leon A. Barmuta, Leah Beesley, Francis J. Burdon, Adriano Caliman, Marcos Callisto, Ian C. Campbell, Bradley J. Cardinale, J. Jesús Casas, Ana M. Chará-Serna, Szymon Ciapała, Eric Chauvet, Jose Checo Colón-Gaud, Aydeé Cornejo, Aaron M. Davis, Monika Degebrodt, Emerson S. Dias, María E. Díaz, Michael M. Douglas, Arturo Elosegi, Andrea C. Encalada, Elvira De Eyto, Ricardo Figueroa, Alexander S. Flecker, André Frainer, Juliana S. França, Erica A. García, Gabriela García, Pavel E. García, Mark O. Gessner, Paul S. Giller, Jesús E. Gómez, Sergio Gómez, José F. Gonçalves Jr., Manuel A. S. Graça, Robert O. Hall Jr., Neusa Hamada, Luiz U. Hepp, Cang Hui, Daichi Imazawa, Tomoya Iwata, Edson S. A. Junior, Samuel K. Kariuki, Andrea Landeira-Dabarca, María Leal, Kaisa Lehosmaa, Charles M. M’ Erimba, Richard Marchant, Renato T. Martins, Frank O. Masese, Megan Camden, Brendan G. Mckie, Adriana O. Medeiros, Jen A. Middleton, Timo Muotka, Junjiro N. Negishi, Jesús Pozo, Alonso Ramírez, Renan S. Rezende, John S. Richardson, José Rincón, Juan Rubio-Ríos, Claudia Serrano, Angela R. Shaffer, Fran Sheldon, Christopher M. Swan, Nathalie S. D. Tenkiano, Scott D. Tiegs, Janine R. Tolod, Michael Vernasky, Anne Watson, Mourine J. Yegon, Catherine M. Yule

Biology: Faculty Publications

Running waters contribute substantially to global carbon fluxes through decomposition of terrestrial plant litter by aquatic microorganisms and detritivores. Diversity of this litter may influence instream decomposition globally in ways that are not yet understood. We investigated latitudinal differences in decomposition of litter mixtures of low and high functional diversity in 40 streams on 6 continents and spanning 113° of latitude. Despite important variability in our dataset, we found latitudinal differences in the effect of litter functional diversity on decomposition, which we explained as evolutionary adaptations of litter-consuming detritivores to resource availability. Specifically, a balanced diet effect appears to operate …

Serial Block-Face Scanning Electron Microscopy (Sbf-Sem) Of Biological Tissue Samples, Justin A Courson, Paul T Landry, Thao Do, Eric Spehlmann, Pascal J Lafontant, Nimesh Patel, Rolando E Rumbaut, Alan R Burns

Faculty, Staff and Students Publications

Serial block-face scanning electron microscopy (SBF-SEM) allows for the collection of hundreds to thousands of serially-registered ultrastructural images, offering an unprecedented three-dimensional view of tissue microanatomy. While SBF-SEM has seen an exponential increase in use in recent years, technical aspects such as proper tissue preparation and imaging parameters are paramount for the success of this imaging modality. This imaging system benefits from the automated nature of the device, allowing one to leave the microscope unattended during the imaging process, with the automated collection of hundreds of images possible in a single day. However, without appropriate tissue preparation cellular ultrastructure can …

A Comparison Of Intertidal Metazoan Biodiversity Between Previously Oiled Sheared And Intact Marsh Margins And Between Multiple Salinity Zones In The Coastal Marshes Of Louisiana, Patrick M. Rayle

LSU Master's Theses

Marshes in Louisiana are under threat from numerous natural and anthropogenic sources. A consequence of these threats are sheared marsh margins, which result from the impact of storm surge on previously oiled, weakened marsh. These conditions occurred in Louisiana marshes after Hurricane Isaac in 2012 followed the Deepwater Horizon oil spill in 2010, particularly in the shorelines surrounding Bay Jimmy. The second and third chapters of this thesis focus on the differences in biodiversity between the sheared and intact marsh margins in impacted sites in Bay Jimmy. Metabarcoding methods were used to determine community composition of the sediment within marsh …

Non-Coding Rna Influence On Manganese Peroxidase Quantity Variation In Isogenic Corn Lines., Amber Booth

Student Theses and Dissertations

New technologies are vital for growing industries that require large amounts of enzymes at a cheaper production cost. Plant transformation technologies coupled with established cropping systems have allowed us to produce enzymes at more efficient rates than ever before. During breeding, offspring are selected for high production of the enzyme of interest and discontinues lines of low production. This continued range is an observed phenomenon in increasingly isogenic lines, but not fully understood. We hypothesize that distinct populations of non-coding RNAs in isogenic corn lines influence enzyme accumulation in seed. This project sequenced high and low manganese peroxidase (MnP)-expressing families …

A Computational Investigation Of The Biophysical Mechanisms Underlying Thermotaxis In The Afd Neurons Of Caenorhabditis Elegans, Zachary Mobille

Theses and Dissertations

Thermotaxis in the nematode Caenorhabditis elegans (C. elegans) is studied at the cellular scale of the amphid finger-like ciliated (AFD) neurons, which have previously been shown to be essential for thermoreception. The voltage and calcium signals of AFD during temperature stimuli are described with ordinary differential equations. The primary calcium model is a modified version of that published by Kuramochi and Doi in 2017 to explain the calcium responses of the chemosensitive amphid single-ciliated right (ASER) neuron to fluctuations in extracellular salt concentration. To account for the effects of temperature, changes to the stimuli conditions under which inactivation takes place …

Identification Of Individual Components Of A Commercial Wheat Germ Acid Phosphatase Preparation, Veronica Moorman, Alexandra Brayton

Chemistry & Biochemistry Publications

Wheat germ acid phosphatase (WGAP) is a commercial preparation of partially purified protein commonly used in laboratory settings for non-specific enzymatic dephosphorylation. It is known that these preparations contain multiple phosphatase isozymes and are still relatively crude. This study therefore aimed to identify the protein components of a commercial preparation of wheat germ acid phosphatase using mass spectroscopy and comparative genomics. After one post-purchase purification step, the most prevalent fifteen proteins in the mixture included heat shock proteins, beta-amylases, glucoseribitol dehydrogenases, enolases, and an aminopeptidase. While not among the most abundant components, eight unique dephosphorylation enzymes were also present including …

A Principal Components Analysis Of Factors Associated With Successful Implementation Of An Lvad Decision Support Tool, Kristin M Kostick, Meredith Trejo, Arvind Bhimaraj, Andrew Civitello, Jonathan Grinstein, Douglas Horstmanshof, Ulrich P Jorde, Matthias Loebe, Mandeep R Mehra, Nasir Z Sulemanjee, Vinay Thohan, Barry H Trachtenberg, Nir Uriel, Robert J Volk, Jerry D Estep, J S Blumenthal-Barby

Faculty, Staff and Students Publications

BACKGROUND: A central goal among researchers and policy makers seeking to implement clinical interventions is to identify key facilitators and barriers that contribute to implementation success. Despite calls from a number of scholars, empirical insights into the complex structural and cultural predictors of why decision aids (DAs) become routinely embedded in health care settings remains limited and highly variable across implementation contexts.

METHODS: We examined associations between "reach", a widely used indicator (from the RE-AIM model) of implementation success, and multi-level site characteristics of nine LVAD clinics engaged over 18 months in implementation and dissemination of a decision aid for …

Telsam-Target Protein Fusions Can Form Diffraction-Quality Crystals Without Direct Inter-Polymer Contacts, Moriah Longhurst

Undergraduate Honors Theses

X-ray diffraction is a robust method for determining the detailed 3D structures of specific proteins. However, this requires the formation of well-ordered protein crystals, a process that is time-consuming, expensive, and only has about a 10-30% success rate. New methods are needed to enable the efficient crystallization of challenging proteins. One such technique is explored here, which utilizes a protein polymer (the sterile alpha motif domain of the human protein translocation Ets leukemia, or TELSAM) as a crystallization chaperone to form a more ordered crystal lattice of target proteins and drive crystallization. This method was successfully used to crystallize, collect …

Highlighting Membrane Protein Structure And Function: A Celebration Of The Protein Data Bank, Fei Li, Pascal F. Egea, Alex J. Vecchio, Ignacio Asial, Meghna Gupta, Joana Paulino, Ruchika Bajaj, Miles Sasha Dickinson, Shelagh Fergunson-Miller, Brian C. Monk, Robert M. Stroud

Department of Biochemistry: Faculty Publications

Biological membranes define the boundaries of cells and compartmentalize the chemical and physical processes required for life. Many biological processes are carried out by proteins embedded in or associated with such membranes. Determination of membrane protein (MP) structures at atomic or near-atomic resolution plays a vital role in elucidating their structural and functional impact in biology. This endeavor has determined 1198 unique MP structures as of early 2021. The value of these structures is expanded greatly by deposition of their three-dimensional (3D) coordinates into the Protein Data Bank (PDB) after the first atomic MP structure was elucidated in 1985. Since …

Investigations Of The Structure And Protein-Protein Interactions Of Chlamydia Trachomatis Scc4, Thilini Oshadhi Senarath Ukwaththage

LSU Doctoral Dissertations

Chlamydia trachomatis (CT) is the most common, sexually transmitted bacterial disease (STD) in the world. In the developmental cycle of CT, specific chlamydia chaperone 4 (Scc4) is a unique protein with essential and multiple roles. Hence, Scc4 is significant as a virulence target for therapeutic approaches to treat chlamydial infections. A novel approach was discovered to purify tag free Scc4 by utilizing a 6X-histidine-tag on Scc1 in the co-expressed Scc4:Scc1 complex by capturing the complex on nickel-charged immobilized metal affinity chromatography resin, followed by dissociation of Scc4 with sarkosyl. Using triple resonance NMR experiments, backbone and sidechain resonances …