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Biochemistry, Biophysics and Molecular Biology Publications

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

Gene Pyramids And The Balancing Act Of Keeping Pests At Bay, Gustavo C. Macintosh Sep 2019

Gene Pyramids And The Balancing Act Of Keeping Pests At Bay, Gustavo C. Macintosh

Biochemistry, Biophysics and Molecular Biology Publications

Pyramiding R genes is a common strategy used by breeders to enhance resistance and increase durability of resistance in crops. However, the molecular mechanisms that mediate R gene interactions are not known. Kamphuis et al. (2019) analyzed Medicago truncatula plants carrying two genes that confer resistance to bluegreen aphids. They identified a potential phytohormone crosstalk triggered by the combined R gene action in response to aphid feeding that enhances resistance and minimizes R gene-associated fitness costs to the plant.


Induction Of Ethylene Inhibits Development Of Soybean Sudden Death Syndrome By Inducing Defense-Related Genes And Reducing Fusarium Virguliforme Growth, Noor A. Abdelsamad, Gustavo C. Macintosh, Leonor F.S. Leandro May 2019

Induction Of Ethylene Inhibits Development Of Soybean Sudden Death Syndrome By Inducing Defense-Related Genes And Reducing Fusarium Virguliforme Growth, Noor A. Abdelsamad, Gustavo C. Macintosh, Leonor F.S. Leandro

Biochemistry, Biophysics and Molecular Biology Publications

Ethylene is a gaseous hormone that regulates plant responses to biotic and abiotic stresses. To investigate the importance of ethylene in soybean resistance to Fusarium virguliforme (Fv), the causal agent of sudden death syndrome (SDS), soybean cultivars Williams 82 (SDS-susceptible) and MN1606 (SDS-resistant) were treated 24 h before and 24h after Fv inoculation with either ethephon (ethylene inducer), cobalt chloride (ethylene biosynthesis inhibitor), or 1-MCP (ethylene perception inhibitor). Inoculated plants were grown for 21 days at 24°C in the greenhouse and then evaluated for SDS severity and expression of soybean defense genes. In both cultivars, plants treated with ethephon ...


Aptamers For Diagnostics With Applications For Infectious Diseases, Muslum Ilgu, Rezzan Fazlioglu, Meric Ozturk, Yasemin Ozsurekci, Marit Nilsen-Hamilton Apr 2019

Aptamers For Diagnostics With Applications For Infectious Diseases, Muslum Ilgu, Rezzan Fazlioglu, Meric Ozturk, Yasemin Ozsurekci, Marit Nilsen-Hamilton

Biochemistry, Biophysics and Molecular Biology Publications

Aptamers are in vitro selected oligonucleotides (DNA, RNA, oligos with modified nucleotides) that can have high affinity and specificity for a broad range of potential targets with high affinity and specificity. Here we focus on their applications as biosensors in the diagnostic field, although they can also be used as therapeutic agents. A small number of peptide aptamers have also been identified. In analytical settings, aptamers have the potential to extend the limit of current techniques as they offer many advantages over antibodies and can be used for real-time biomarker detection, cancer clinical testing, and detection of infectious microorganisms and ...


Multiple Variables At The Leukocyte Cell Surface Impact Fc Γ Receptor-Dependent Mechanisms, Kashyap R. Patel, Jacob T. Roberts, Adam W. Barb Feb 2019

Multiple Variables At The Leukocyte Cell Surface Impact Fc Γ Receptor-Dependent Mechanisms, Kashyap R. Patel, Jacob T. Roberts, Adam W. Barb

Biochemistry, Biophysics and Molecular Biology Publications

Fc γ receptors (FcγR) expressed on the surface of human leukocytes bind clusters of immunoglobulin G (IgG) to induce a variety of responses. Many therapeutic antibodies and vaccine-elicited antibodies prevent or treat infectious diseases, cancers and autoimmune disorders by binding FcγRs, thus there is a need to fully define the variables that impact antibody-induced mechanisms to properly evaluate candidate therapies and design new intervention strategies. A multitude of factors influence the IgG-FcγR interaction; one well-described factor is the differential affinity of the six distinct FcγRs for the four human IgG subclasses. However, there are several other recently described factors that ...


Entropy, Fluctuations, And Disordered Proteins, Eshel Faraggi, A. Keith Dunker, Robert L. Jernigan, Andrzej Kloczkowski Jan 2019

Entropy, Fluctuations, And Disordered Proteins, Eshel Faraggi, A. Keith Dunker, Robert L. Jernigan, Andrzej Kloczkowski

Biochemistry, Biophysics and Molecular Biology Publications

Entropy should directly reflect the extent of disorder in proteins. By clustering structurally related proteins and studying the multiple-sequence-alignment of the sequences of these clusters, we were able to link between sequence, structure, and disorder information. We introduced several parameters as measures of fluctuations at a given MSA site and used these as representative of the sequence and structure entropy at that site. In general, we found a tendency for negative correlations between disorder and structure, and significant positive correlations between disorder and the fluctuations in the system. We also found evidence for residue-type conservation for those residues proximate to ...


Switching On A Nontraditional Enzymatic Base—Deprotonation By Serine In The Ent-Kaurene Synthase From Bradyrhizobium Japonicum, Meirong Jia, Yue Zhang, Justin B. Siegel, Dean J. Tantillo, Reuben J. Peters Jan 2019

Switching On A Nontraditional Enzymatic Base—Deprotonation By Serine In The Ent-Kaurene Synthase From Bradyrhizobium Japonicum, Meirong Jia, Yue Zhang, Justin B. Siegel, Dean J. Tantillo, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Terpene synthases often catalyze complex carbocation cascade reactions. It has been previously shown that single-residue switches involving replacement of a key aliphatic residue with serine or threonine can “short-circuit” such reactions that are presumed to act indirectly via dipole stabilization of intermediate carbocations. Here a similar switch was found in the structurally characterized ent-kaurene synthase from Bradyrhizobium japonicum. Application of a recently developed computational approach to terpene synthases, TerDockin, surprisingly indicates direct action of the introduced serine hydroxyl as a catalytic base. Notably, this model suggests an alternative interpretation of previous results and potential routes toward reengineering terpene synthase ...


A Single-Cell Platform For Reconstituting And Characterizing Fatty Acid Elongase Component Enzymes, Alexis A. Campbell, Kenna E. Stenback, Kayla Flyckt, Trang Hoang, M. Ann Perera, Basil J. Nikolau Jan 2019

A Single-Cell Platform For Reconstituting And Characterizing Fatty Acid Elongase Component Enzymes, Alexis A. Campbell, Kenna E. Stenback, Kayla Flyckt, Trang Hoang, M. Ann Perera, Basil J. Nikolau

Biochemistry, Biophysics and Molecular Biology Publications

Fatty acids of more than 18-carbons, generally known as very long chain fatty acids (VLCFAs) are essential for eukaryotic cell viability, and uniquely in terrestrial plants they are the precursors of the cuticular lipids that form the organism’s outer barrier to the environment. VLCFAs are synthesized by fatty acid elongase (FAE), which is an integral membrane enzyme system with multiple components. The genetic complexity of the FAE system, and its membrane association has hampered the biochemical characterization of FAE. In this study we computationally identified Zea mays genetic sequences that encode the enzymatic components of FAE and developed a ...


Evidence For A Role Of Spindle Matrix Formation In Cell Cycle Progression By Antibody Perturbation, Changfu Yao, Chao Wang, Yeran Li, Michael Zavortink, Vincent Archambault, Jack Girton, Kristen M. Johansen, Jorgen Johansen Nov 2018

Evidence For A Role Of Spindle Matrix Formation In Cell Cycle Progression By Antibody Perturbation, Changfu Yao, Chao Wang, Yeran Li, Michael Zavortink, Vincent Archambault, Jack Girton, Kristen M. Johansen, Jorgen Johansen

Biochemistry, Biophysics and Molecular Biology Publications

In Drosophila it has recently been demonstrated that a spindle matrix in the form of a membrane-less macromolecular assembly embeds the microtubule-based spindle apparatus. In addition, two of its constituents, Megator and Chromator, were shown to function as spatial regulators of spindle checkpoint proteins. However, whether the spindle matrix plays a wider functional role in spatially regulating cell cycle progression factors was unknown. Here using a live imaging approach we provide evidence that a number of key cell cycle proteins such as Cyclin B, Polo, and Ran co-localize with the spindle matrix during mitosis. Furthermore, prevention of spindle matrix formation ...


A Third Class: Functional Gibberellin Biosynthetic Operon In Beta-Proteobacteria, Raimund Nagel, John E. Bieber, Mark G. Schmidt-Dannert, Ryan S. Nett, Reuben J. Peters Nov 2018

A Third Class: Functional Gibberellin Biosynthetic Operon In Beta-Proteobacteria, Raimund Nagel, John E. Bieber, Mark G. Schmidt-Dannert, Ryan S. Nett, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

The ability of plant-associated microbes to produce gibberellin A (GA) phytohormones was first described for the fungal rice pathogen Gibberella fujikuroi in the 1930s. Recently the capacity to produce GAs was shown for several bacteria, including symbiotic alpha-proteobacteria (α-rhizobia) and gamma-proteobacteria phytopathogens. All necessary enzymes for GA production are encoded by a conserved operon, which appears to have undergone horizontal transfer between and within these two phylogenetic classes of bacteria. Here the operon was shown to be present and functional in a third class, the beta-proteobacteria, where it is found in several symbionts (β-rhizobia). Conservation of function was examined by ...


Arginine In The Farm And Sarm: A Role In Chain-Length Determination For Arginine In The Aspartate-Rich Motifs Of Isoprenyl Diphosphate Synthases From Mycobacterium Tuberculosis, Raimund Nagel, Jill A. Thomas, Faith A. Adekunle, Francis M. Mann, Reuben J. Peters Oct 2018

Arginine In The Farm And Sarm: A Role In Chain-Length Determination For Arginine In The Aspartate-Rich Motifs Of Isoprenyl Diphosphate Synthases From Mycobacterium Tuberculosis, Raimund Nagel, Jill A. Thomas, Faith A. Adekunle, Francis M. Mann, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Isoprenyl chains are found in many important metabolites. These are derived from precursors of the appropriate length produced by isoprenyl diphosphate synthases (IDSs). The human pathogen Mycobacterium tuberculosis makes various isoprenoids/terpenoids, with important roles in their biosynthesis played by two closely related IDSs, encoded by grcC1 (Rv0562) and grcC2 (Rv0989c), with Rv0989c generating the 10-carbon precursor (E)-geranyl diphosphate (GPP), and Rv0562 the 20-carbon precursor (E,E,E)-geranylgeranyl diphosphate (GGPP). Intriguingly, while Rv0562 contains the prototypical trans-IDS first and second aspartate-rich (DDxxD) motifs (FARM and SARM, respectively), Rv0989c uniquely contains arginine in place of the second Asp in ...


Continuous Interdomain Orientation Distributions Reveal Components Of Binding Thermodynamics, Yang Qi, Jeffrey W. Martin, Adam W. Barb, Francois Thelot, Anthony Yan, Bruce R. Donald, Terrence G. Oas Sep 2018

Continuous Interdomain Orientation Distributions Reveal Components Of Binding Thermodynamics, Yang Qi, Jeffrey W. Martin, Adam W. Barb, Francois Thelot, Anthony Yan, Bruce R. Donald, Terrence G. Oas

Biochemistry, Biophysics and Molecular Biology Publications

The flexibility of biological macromolecules is an important structural determinant of function. Unfortunately, the correlations between different motional modes are poorly captured by discrete ensemble representations. Here, we present new ways to both represent and visualize correlated interdomain motions. Interdomain motions are determined directly from residual dipolar couplings, represented as a continuous conformational distribution, and visualized using the disk-on-sphere representation. Using the disk-on-sphere representation, features of interdomain motions, including correlations, are intuitively visualized. The representation works especially well for multidomain systems with broad conformational distributions.This analysis also can be extended to multiple probability density modes, using a Bingham mixture ...


A 2′Fy-Rna Motif Defines An Aptamer For Ebolavirus Secreted Protein, Shambhavi Shubham, Jan Hoinka, Soma Banerjee, Emma Swanson, Jacob A. Dillard, Nicholas J. Lennemann, Teresa M. Przytycka, Wendy Maury, Marit Nilsen-Hamilton Aug 2018

A 2′Fy-Rna Motif Defines An Aptamer For Ebolavirus Secreted Protein, Shambhavi Shubham, Jan Hoinka, Soma Banerjee, Emma Swanson, Jacob A. Dillard, Nicholas J. Lennemann, Teresa M. Przytycka, Wendy Maury, Marit Nilsen-Hamilton

Biochemistry, Biophysics and Molecular Biology Publications

With properties such as stability to long-term storage and amenability to repetitive use, nucleic acid aptamers are compatible with many sensing/transducing platforms intended for use in remote locations. Sensors with these properties are important for quickly identifying ebolavirus outbreaks, which frequently start in locations that lack sophisticated equipment. Soluble glycoprotein (sGP), an excellent biomarker for ebolaviruses, is produced from the same gene as the ebolavirus glycoprotein GP1,2 that decorates the surface of the viral particle and is secreted in abundance into the blood stream even during the early stages of infection. Here, we report the selection and properties ...


Direct Determination Of The Site Of Addition Of Glucosyl Units To Maltooligosaccharide Acceptors Catalyzed By Maize Starch Synthase I, Ying Xie, Adam W. Barb, Tracie A. Hennen-Bierwagen, Alan M. Myers Aug 2018

Direct Determination Of The Site Of Addition Of Glucosyl Units To Maltooligosaccharide Acceptors Catalyzed By Maize Starch Synthase I, Ying Xie, Adam W. Barb, Tracie A. Hennen-Bierwagen, Alan M. Myers

Biochemistry, Biophysics and Molecular Biology Publications

Starch synthase (SS) (ADP-glucose:1,4-α-D-glucan 4-α-D-glucosyltransferase) elongates α-(1→4)-linked linear glucans within plastids to generate the storage polymers that constitute starch granules. Multiple SS classes are conserved throughout the plant kingdom, indicating that each provides a unique function responsible for evolutionary selection. Evidence has been presented arguing for addition of glucosyl units from the ADPglucose donor to either the reducing end or the non-reducing end of the acceptor substrate, although until recently direct evidence addressing this question was not available. Characterization of newly incorporated glucosyl units determined that recombinant maize (Zea mays L.) SSIIa elongates its substrates ...


Protein Dynamic Communities From Elastic Network Models Align Closely To The Communities Defined By Molecular Dynamics, Sambit Kumar Mishra, Robert L. Jernigan Jun 2018

Protein Dynamic Communities From Elastic Network Models Align Closely To The Communities Defined By Molecular Dynamics, Sambit Kumar Mishra, Robert L. Jernigan

Biochemistry, Biophysics and Molecular Biology Publications

Dynamic communities in proteins comprise the cohesive structural units that individually exhibit rigid body motions. These can correspond to structural domains, but are usually smaller parts that move with respect to one another in a protein’s internal motions, key to its functional dynamics. Previous studies emphasized their importance to understand the nature of ligand-induced allosteric regulation. These studies reported that mutations to key community residues can hinder transmission of allosteric signals among the communities. Usually molecular dynamic (MD) simulations (~ 100 ns or longer) have been used to identify the communities—a demanding task for larger proteins. In the present ...


Probing The Specificity Of Cyp112 In Bacterial Gibberellin Biosynthesis, Raimund Nagel, Reuben J. Peters Jun 2018

Probing The Specificity Of Cyp112 In Bacterial Gibberellin Biosynthesis, Raimund Nagel, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Biosynthesis of the gibberellin A (GA) plant hormones evolved independently in plant-associated fungi and bacteria. While the relevant enzymes have distinct evolutionary origins, the pathways proceed via highly similar reactions. One particularly complex transformation involves combined demethylation and γ-lactone ring formation, catalyzed in bacteria by the cytochrome P450 CYP112 in three individual steps, which involves large structural changes in the transition from substrate to product, with further divergence in the recently demonstrated use of two separate mechanistic routes. Here the substrate specificity of the isozyme from Erwinia tracheiphila, EtCYP112, was probed via UV-Vis spectral binding studies and activity assays with ...


Direct Production Of Dihydroxylated Sesquiterpenoids By A Maize Terpene Synthase, Jin Liang, Jiang Liu, Reid Brown, Meirong Jia, Ke Zhou, Reuben J. Peters, Qiang Wang Jun 2018

Direct Production Of Dihydroxylated Sesquiterpenoids By A Maize Terpene Synthase, Jin Liang, Jiang Liu, Reid Brown, Meirong Jia, Ke Zhou, Reuben J. Peters, Qiang Wang

Biochemistry, Biophysics and Molecular Biology Publications

The astounding structural and biological diversity of the large class of terpenoid natural products are imparted by both their complex hydrocarbon backbones and further elaboration by the addition of multiple hydroxyl groups, which provide both solubility and specific binding properties. While the role of terpene synthases in generating hydrocarbons with complex backbones is well known, these also are known to generate (singly) hydroxylated products by the addition of water prior to terminating deprotonation. Here a maize sesquiterpene synthase was unexpectedly found to generate dually hydroxylated products directly from (E,E)-farnesyl diphosphate, primarily eudesmane-2,11-diol, along with two closely related ...


A Dendritic Guidance Receptor Complex Brings Together Distinct Actin Regulators To Drive Efficient F-Actin Assembly And Branching, Wei Zhou, Xintong Dong, Timothy R. Broederdorf, Ao Shen, Daniel A. Kramer, Rebecca Shi, Xing Liang, David M. Miller Iii, Yang K. Xiang, Ryohei Yasuda, Baoyu Chen, Kang Shen May 2018

A Dendritic Guidance Receptor Complex Brings Together Distinct Actin Regulators To Drive Efficient F-Actin Assembly And Branching, Wei Zhou, Xintong Dong, Timothy R. Broederdorf, Ao Shen, Daniel A. Kramer, Rebecca Shi, Xing Liang, David M. Miller Iii, Yang K. Xiang, Ryohei Yasuda, Baoyu Chen, Kang Shen

Biochemistry, Biophysics and Molecular Biology Publications

Proper morphogenesis of dendrites plays a fundamental role in the establishment of neural circuits. The molecular mechanism by which dendrites grow highly complex branches is not well understood. Here, using the C. elegans PVD neuron, we demonstrate that high-order dendritic branching requires actin polymerization driven by coordinated interactions between two membrane proteins, DMA-1 and HPO-30, and with their cytoplasmic interactors, the RacGEF TIAM-1 and the actin nucleation promotion factor WAVE Regulatory Complex (WRC). The dendrite branching receptor DMA-1 directly binds to the PDZ domain of TIAM-1, while the claudin-like protein HPO-30 directly interacts with the WRC. On dendrites, DMA-1 and ...


Inferring Roles In Defense From Metabolic Allocation Of Rice Diterpenoids, Xuan Lu, Juan Zhang, Benjamin Brown, Riqing Li, Julio Rodríguez-Romero, Aileen Berasategui, Bo Liu, Meimei Xu, Dangping Luo, Zhiqiang Pan, Scott R. Baerson, Jonathan Gershenzon, Zhaohu Li, Ane Sesma, Bing Yang, Reuben J. Peters May 2018

Inferring Roles In Defense From Metabolic Allocation Of Rice Diterpenoids, Xuan Lu, Juan Zhang, Benjamin Brown, Riqing Li, Julio Rodríguez-Romero, Aileen Berasategui, Bo Liu, Meimei Xu, Dangping Luo, Zhiqiang Pan, Scott R. Baerson, Jonathan Gershenzon, Zhaohu Li, Ane Sesma, Bing Yang, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Among their responses to microbial infection, plants deploy an arsenal of natural antibiotic products. Historically these have been identified on the basis of their antibiotic activity in vitro, which leaves open the question of their relevance to defense in planta. The vast majority of such natural products from the important crop plant rice (Oryza sativa) are diterpenoids whose biosynthesis proceeds via either ent- or syn-copalyl diphosphate (CPP) intermediates, which were isolated on the basis of their antibiotic activity against the fungal blast pathogen Magnaporthe oryzae. However, rice plants in which the gene for the syn-CPP synthase Os-CPS4 is ...


Restricted Processing Of Cd16a/Fc Γ Receptor Iiia N-Glycans From Primary Human Nk Cells Impacts Structure And Function, Kashyap R. Patel, Jacob T. Roberts, Ganesh P. Subedi, Adam W. Barb Mar 2018

Restricted Processing Of Cd16a/Fc Γ Receptor Iiia N-Glycans From Primary Human Nk Cells Impacts Structure And Function, Kashyap R. Patel, Jacob T. Roberts, Ganesh P. Subedi, Adam W. Barb

Biochemistry, Biophysics and Molecular Biology Publications

CD16a/Fc γ receptor IIIa is the most abundant antibody Fc receptor expressed on human natural killer (NK) cells and activates a protective cytotoxic response following engagement with antibody clustered on the surface of a pathogen or diseased tissue. Therapeutic monoclonal antibodies (mAbs) with greater Fc-mediated affinity for CD16a show superior therapeutic outcome; however, one significant factor that promotes antibody–CD16a interactions, the asparagine-linked carbohydrates (N-glycans), remains undefined. Here, we purified CD16a from the primary NK cells of three donors and identified a large proportion of hybrid (22%) and oligomannose N-glycans (23%). These proportions indicated restricted N-glycan ...


Seqstruct : A New Amino Acid Similarity Matrix Based On Sequence Correlations And Structural Contacts Yields Sequence-Structure Congruence, Kejue Jia, Robert L. Jernigan Feb 2018

Seqstruct : A New Amino Acid Similarity Matrix Based On Sequence Correlations And Structural Contacts Yields Sequence-Structure Congruence, Kejue Jia, Robert L. Jernigan

Biochemistry, Biophysics and Molecular Biology Publications

Protein sequence matching does not properly account for some well-known features of protein structures: surface residues being more variable than core residues, the high packing densities in globular proteins, and does not yield good matches of sequences of many proteins known to be close structural relatives. There are now abundant protein sequences and structures to enable major improvements to sequence matching. Here, we utilize structural frameworks to mount the observed correlated sequences to identify the most important correlated parts. The rationale is that protein structures provide the important physical framework for improving sequence matching. Combining the sequence and structure data ...


Mouse Igg2c Fc Loop Residues Promote Greater Receptor-Binding Affinity Than Mouse Igg2b Or Human Igg1, Daniel J. Falconer, Adam W. Barb Feb 2018

Mouse Igg2c Fc Loop Residues Promote Greater Receptor-Binding Affinity Than Mouse Igg2b Or Human Igg1, Daniel J. Falconer, Adam W. Barb

Biochemistry, Biophysics and Molecular Biology Publications

The structures of non-human antibodies are largely unstudied despite the potential for the identification of alternative structural motifs and physical properties that will benefit a basic understanding of protein and immune system evolution as well as highlight unexplored motifs to improve therapeutic monoclonal antibody. Here we probe the structure and receptor-binding properties of the mouse IgG2c crystallizable fragment (Fc) to compare to mouse IgG2b and human IgG1 Fcs. Models of mIgG2c Fc determined by x-ray crystallography with a complex-type biantennary (to 2.05 Å) or a truncated (1)GlcNAc asparagine-linked (N)-glycan attached (to 2.04 Å) show differences in ...


Comparisons Of Protein Dynamics From Experimental Structure Ensembles, Molecular Dynamics Ensembles, And Coarse-Grained Elastic Network Models, Kannan Sankar, Sambit K. Mishra, Robert L. Jernigan Jan 2018

Comparisons Of Protein Dynamics From Experimental Structure Ensembles, Molecular Dynamics Ensembles, And Coarse-Grained Elastic Network Models, Kannan Sankar, Sambit K. Mishra, Robert L. Jernigan

Biochemistry, Biophysics and Molecular Biology Publications

Predicting protein motions is important for bridging the gap between protein structure and function. With growing numbers of structures of the same, or closely related proteins becoming available, it is now possible to understand more about the intrinsic dynamics of a protein with principal component analysis (PCA) of the motions apparent within ensembles of experimental structures. In this paper, we compare the motions extracted from experimental ensembles of 50 different proteins with the modes of motion predicted by several types of coarse-grained elastic network models (ENMs) which additionally take into account more details of either the protein geometry or the ...


Aptamer-Enabled Uptake Of Small Molecule Ligands, Supipi Liyamali Auwardt, Yeon-Jung Seo, Muslum Ilgu, Judhajeet Ray, Robert R. Feldges, Shambhavi Shubham, Lee Bendickson, Howard A. Levine, Marit Nilsen-Hamilton Jan 2018

Aptamer-Enabled Uptake Of Small Molecule Ligands, Supipi Liyamali Auwardt, Yeon-Jung Seo, Muslum Ilgu, Judhajeet Ray, Robert R. Feldges, Shambhavi Shubham, Lee Bendickson, Howard A. Levine, Marit Nilsen-Hamilton

Biochemistry, Biophysics and Molecular Biology Publications

The relative ease of isolating aptamers with high specificity for target molecules suggests that molecular recognition may be common in the folds of natural RNAs. We show here that, when expressed in cells, aptamers can increase the intracellular concentrations of their small molecule ligands. We have named these aptamers as DRAGINs (Drug Binding Aptamers for Growing Intracellular Numbers). The DRAGIN property, assessed here by the ability to enhance the toxicity of their ligands, was found for some, but not all, aminoglycoside aptamers. One aptamer protected cells against killing by its ligand. Another aptamer promoted killing as a singlemer and protected ...


Biosynthesis Of Diterpenoids In Tripterygium Adventitious Root Cultures, Fainmarinat S. Inabuy, Justin T. Fischedick, Iris Lange, Michael Hartmann, Narayanan Srividya, Amber N. Parrish, Meimei Xu, Reuben J. Peters, B. Markus Lange Sep 2017

Biosynthesis Of Diterpenoids In Tripterygium Adventitious Root Cultures, Fainmarinat S. Inabuy, Justin T. Fischedick, Iris Lange, Michael Hartmann, Narayanan Srividya, Amber N. Parrish, Meimei Xu, Reuben J. Peters, B. Markus Lange

Biochemistry, Biophysics and Molecular Biology Publications

Adventitious root cultures were developed from Tripterygium regelii Sprague & Takeda and growth conditions optimized for the abundant production of diterpenoids, which can be collected directly from the medium. An analysis of publicly available transcriptome data sets collected with T. regelii roots and root cultures indicated the presence of a large gene family (with 20 members) for terpene synthases (TPSs). Nine candidate diterpene synthase genes were selected for follow-up functional evaluation, of which two belonged to the TPS-c, three to the TPS-e/f and four to the TPS-b subfamily. These genes were characterized by heterologous expression in a modular metabolic engineering ...


Carbohydrate–Polypeptide Contacts In The Antibody Receptor Cd16a Identified Through Solution Nmr Spectroscopy, Ganesh P. Subedi, Daniel J. Falconer, Adam W. Barb Jun 2017

Carbohydrate–Polypeptide Contacts In The Antibody Receptor Cd16a Identified Through Solution Nmr Spectroscopy, Ganesh P. Subedi, Daniel J. Falconer, Adam W. Barb

Biochemistry, Biophysics and Molecular Biology Publications

Asparagine-linked carbohydrates (N-glycans) are a common modification of eukaryotic proteins that confer multiple properties including the essential stabilization of therapeutic monoclonal antibodies. Here we present a rapid and efficient strategy to identify N-glycans that contact polypeptide residues and apply the method to profile the five N-glycans attached to the human antibody receptor CD16A (Fc γ receptor IIIA). Human embryonic kidney 293S cells expressed CD16A with [13CU]-labeled N-glycans using standard protein expression techniques and medium supplemented with 3 g/L [13CU]-glucose. Anomeric resonances on the protein-linked N-acetylglucosamine residue at the reducing end of the glycan are particularly well suited ...


Investigating The Phylogenetic Range Of Gibberellin Biosynthesis In Bacteria, Raimund Nagel, Reuben J. Peters Apr 2017

Investigating The Phylogenetic Range Of Gibberellin Biosynthesis In Bacteria, Raimund Nagel, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Certain plant-associated microbes can produce gibberellin (GA) phytohormones, as first described for the rice fungal pathogen Gibberella fujikuroi and, more recently, for bacteria, including several rhizobia and the rice bacterial pathogen Xanthomonas oryzae pv. oryzicola. The relevant enzymes are encoded by a biosynthetic operon that exhibits both a greater phylogenetic range and scattered distribution among plant-associated bacteria. Here, the phylogenetic distribution of this operon was investigated. To demonstrate conserved functionality, the enzymes encoded by the disparate operon from X. translucens pv. translucens, along with those from the most divergent example, found in Erwinia tracheiphila, were biochemically characterized. In both of ...


Data On Il-10r Neutralization-Induced Chronic Colitis In Lipocalin 2 Deficient Mice On Balb/C Background, Vishal Singh, Beng San Yeoh, Marit Nilsen-Hamilton, Thorsten Berger, Tak W. Mak, Matam Vijay-Kumar Apr 2017

Data On Il-10r Neutralization-Induced Chronic Colitis In Lipocalin 2 Deficient Mice On Balb/C Background, Vishal Singh, Beng San Yeoh, Marit Nilsen-Hamilton, Thorsten Berger, Tak W. Mak, Matam Vijay-Kumar

Biochemistry, Biophysics and Molecular Biology Publications

The data herein is related to the research article entitled “Microbiota-inducible Innate Immune, Siderophore Binding Protein Lipocalin 2 is Critical for Intestinal Homeostasis” (Singh et al., 2016) [1] where we have demonstrated that C57BL/6 Lipocalin 2 deficient mice (Lcn2KO) developed chronic colitis upon anti-interleukin-10 receptor (αIL-10R) monoclonal antibody administration. In the present article, we evaluated the susceptibility of BALB/c Lcn2KO mice and their WT littermates to the αIL-10R neutralization-induced chronic colitis. Our data showed that αIL-10R mAb-treated BALB/c Lcn2KO mice exhibited severe chronic colitis (i.e., splenomegaly, colomegaly, colonic pathology, and incidence of rectal ...


A Pair Of Residues That Interactively Affect Diterpene Synthase Product Outcome, Meirong Jia, Samuel Tufts, Samuel Schulte, Reuben J. Peters Jan 2017

A Pair Of Residues That Interactively Affect Diterpene Synthase Product Outcome, Meirong Jia, Samuel Tufts, Samuel Schulte, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

The labdane-related diterpenoids (LRDs) are an important superfamily of natural products whose structural diversity critically depends on the hydrocarbon skeletal structures generated, in large part, by class I diterpene synthases. In the plant kingdom, where the LRDs are predominantly found, the relevant class I diterpene synthases are clearly derived from the ent-kaurene synthase (KS) required in all land plants for phytohormone biosynthesis and, hence, are often termed KS-like (KSL). Previous work, initiated by the distinct function of two alleles of a KSL from rice, OsKSL5, identified a single residue switch with a profound effect on not only OsKSL5 product ...


H2av Facilitates H3s10 Phosphorylation But Is Not Required For Heat Shock-Induced Chromatin Decondensation Or Transcriptional Elongation, Yeran Li, Chao Wang, Weili Cai, Saheli Sengupta, Michael Zavortink, Huai Deng, Jack Girton, Jorgen Johansen, Kristen M. Johansen Jan 2017

H2av Facilitates H3s10 Phosphorylation But Is Not Required For Heat Shock-Induced Chromatin Decondensation Or Transcriptional Elongation, Yeran Li, Chao Wang, Weili Cai, Saheli Sengupta, Michael Zavortink, Huai Deng, Jack Girton, Jorgen Johansen, Kristen M. Johansen

Biochemistry, Biophysics and Molecular Biology Publications

A model has been proposed in which JIL-1 kinase-mediated H3S10 and H2Av phosphorylation is required for transcriptional elongation and heat shock-induced chromatin decondensation. However, here we show that although H3S10 phosphorylation is indeed compromised in the H2Av null mutant, chromatin decondensation at heat shock loci is unaffected in the absence of JIL-1 as well as of H2Av and that there is no discernable decrease in the elongating form of RNA polymerase II in either mutant. Furthermore, mRNA for the major heat shock protein Hsp70 is transcribed at robust levels in both H2Avand JIL-1 null mutants. Using a different chromatin ...


Cis Or Trans With Class Ii Diterpene Cyclases, Meirong Jia, Reuben J. Peters Jan 2017

Cis Or Trans With Class Ii Diterpene Cyclases, Meirong Jia, Reuben J. Peters

Biochemistry, Biophysics and Molecular Biology Publications

Isoprenoid precursors readily undergo (poly)cyclization in electrophilic reaction cascades, presumably as internal addition of the carbon–carbon double-bonds from neighboring isoprenyl repeats readily forms relatively stable cyclohexyl tertiary carbocation intermediates. This hypothesis is agnostic regarding alkene configuration (i.e., Z or E). Consistent with this, here it is shown that certain class II diterpene cyclases, which normally convert (E,E,E)-geranylgeranyl diphosphate to 13E-trans-decalin bicycles, will also act upon (Z,Z,Z)-nerylneryl diphosphate, producing novel 13Z-cis-decalin bicycles instead.