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Tadao Takada, Syunya Ishino, Ami Takata, Mitsunobu Nakamura, Mamoru Fujitsuka, Tetsuro Majima, Kazushige Yamana
We have demonstrated the rapid electron transfer in homo- and hetero-complexes composed of the parent perylenediimide (PDI, PH) and pyrrolidine-substituted PDI (PN) in DNA analyzed by steady-state fluorescence and femtosecond transient absorption measurements. The DNA molecules possessing PH and PN were prepared by our recently developed method using the reaction of enzymatically-generated abasic sites, by which the PDI molecules can be incorporated as base surrogates within the DNA base stack. The melting temperature analysis showed that the PH and PN monomer, and their homo- and heterodimers can contribute to the stabilization of the DNA duplex...
April 15, 2018: Chemistry: a European Journal
Yunier Rodríguez-Álvarez, Ania Cabrales-Rico, Alejandro Perera-Pintado, Anais Prats-Capote, Hilda E Garay-Pérez, Osvaldo Reyes-Acosta, Erik Pérez-García, Araceli Chico-Capote, Alicia Santos-Savio
Interleukin (IL)-15 is an inflammatory cytokine that constitutes a validated therapeutic target in some immunopathologies, including rheumatoid arthritis (RA). Previously, we identified an IL-15 antagonist peptide named [K6T]P8, with potential therapeutic application in RA. In the current work, the metabolic stability of this peptide in synovial fluids from RA patients was studied. Moreover, [K6T]P8 peptide was labeled with 99m Tc to investigate its stability in human plasma and its biodistribution pattern in healthy rats...
April 15, 2018: Journal of Peptide Science: An Official Publication of the European Peptide Society
Dmitri Konarev, Alexey Kuzmin, Salavat Khasanov, Aleksey Fatalov, Evgenia Yudanova, Rimma Lyubovskaya
Reduction methods for preparation of coordination complexes of titanium(II) and indium(III) phthalocyanines with organic dyes such as indigo, thioindigo and squarylium dye III (SQ) have been developed which allow one to obtain crystalline {cryptand(K+)}{(cis-indigo-O,O)(0)Ti(II)[Pc(2-)}(Cl-).C6H4Cl2 (1), {cryptand(K+)}{(cis-thioindigo-O, O)(2-)In(III)[Pc(2-)]}(-).C6H4Cl2 (2) and {cryptand(K+)}{[(SQ)2-O, O](2-)In(III)[Pc(2-)]}(-).3.5C6H4Cl2 (3) complexes. The formation of these complexes is accompanied by the reduction of starting dyes or metal phthalocyanines...
April 14, 2018: Chemistry: a European Journal
Blair Alden Parry, Anna Marie Chang, Sebastian M Schellong, Stacey L House, Gregory J Fermann, Erin K Deadmon, Nicholas J Giordano, Yuchiao Chang, Jason Cohen, Nancy Robak, Adam J Singer, Mary Mulrow, Ellen T Reibling, Samuel Francis, S Michelle Griffin, Jürgen H Prochaska, Barbara Davis, Patricia McNelis, Joao Delgado, Philipp Kümpers, Nikos Werner, Nina T Gentile, Eli Zeserson, Philipp S Wild, Alexander T Limkakeng, Elizabeth Lea Walters, Frank LoVecchio, Daniel Theodoro, Judd E Hollander, Christopher Kabrhel
INTRODUCTION: We sought to determine the test characteristics of an automated INNOVANCE D-dimer assay for the exclusion of pulmonary embolism (PE) and deep venous thrombosis (DVT) in emergency department (ED) patients using standard and age-adjusted cut-offs. METHODS: Cross-sectional, international, multicenter study of consecutive patients with suspected DVT or PE in 24 centers (18 USA, 6 Europe). Evaluated patients had low or intermediate Wells PE or DVT scores...
April 5, 2018: Thrombosis Research
Lin-Gao Ju, Xiang Lin, Dong Yan, Qing-Lan Li, Min Wu, Lian-Yun Li
ERAD is an important process of protein quality control that eliminates misfolded or unassembled proteins from ER. Before undergoing proteasome degradation, the misfolded proteins are dislocated from ER membrane into cytosol, which requires the AAA ATPase p97/VCP and its cofactor, the NPL4-UFD1 dimer. Here, we performed a CRISPR-based screen and identify many candidates for ERAD regulation. We further confirmed four proteins, FBOX2, TRIM6, UFL1 and WDR20, are novel regulators for ERAD. Then the molecular mechanism for WDR20 in ERAD is further characterized...
April 12, 2018: Biochimica et Biophysica Acta
Sujeewa D Wijesuriya, Elizabeth Pongo, Milan Tomic, Fangjiu Zhang, Consuelo Garcia-Rodriquez, Fraser Conrad, Shauna Farr-Jones, James D Marks, Arnold H Horwitz
Expression variation among antibodies produced by stably transfected Chinese Hamster Ovary (CHO) cells is well established. While developing CHO-K1 cell lines, we encountered a human monoclonal antibody, mAb B-c, with severe manufacturability issues, including very poor expression and high levels of heavy chain (HC) dimer and high molecular weight aggregates. Using transient expression in CHO-K1 cells, we identified light chain (LC) as the source of the manufacturability issues for this antibody. While other antibodies achieved optimal expression at 1:1 or 2:1 LC to HC ratios, mAb B-c required up to a 6:1 LC:HC for maximal expression, which was still significantly lower than that for other tested antibodies...
April 12, 2018: Protein Expression and Purification
Benjamín Rubio-Jurado, Luz-Ma-Adriana Balderas-Peña, Eduardo E García-Luna, María G Zavala-Cerna, Carlos Riebeling-Navarro, Pedro A Reyes, Arnulfo H Nava-Zavala
Neoplasms exhibits a high incidence and mortality rates due to their complex and commonly overlapping clinical, biochemical, and morphologic profiles influenced by acquired or inherited molecular abnormalities, cell of origin, and level of differentiation. Obesity appears related to ~20% of cancers including endometrial, esophageal, colorectal, postmenopausal breast, prostate, and renal. Several factors other than obesity, i.e., insulin, insulin-like growth factor, sexual hormones, and adipokines may play a potential role in neoplasia...
2018: Advances in Clinical Chemistry
Fu-Hsien Yu, Chin-Tien Wang
BACKGROUND: HIV-1 protease (PR) activation is triggered by Gag-Pol dimerization. Premature PR activation results in reduced virion yields due to enhanced Gag cleavage. A p6* transframe peptide located directly upstream of protease is believed to play a modulating role in PR activation. Previous reports indicate that the C-terminal p6* tetra-peptide prevents premature PR activation triggered by a leucine zipper (LZ) dimerization motif inserted in the deleted p6* region. To clarify the involvement of C-terminal p6* residues in mitigating enhanced LZ-incurred Gag processing, we engineered constructs containing C-terminal p6* residue substitutions with and without a mutation blocking the p6*/PR cleavage site, and created other Gag or p6* domain-removing constructs...
April 14, 2018: Retrovirology
Qing Li, Jason B White, Norman C Peterson, Keith W Rickert, Christopher O Lloyd, Kevin L Allen, Kim Rosenthal, Xizhe Gao, Herren Wu, William F Dall'Acqua, M Jack Borrok, Ping Tsui
The accumulation, dissemination and clearance of monoclonal antibody-based therapeutics or imaging reagents targeting tumor associated antigens is governed by several factors including affinity, size, charge, and valency. Tumor targeting antibody fragments have distinct advantages over intact monoclonal antibodies such as enhanced penetration within the tumor and rapid accumulation but are subject to rapid clearance. Polyethylene glycol (PEG)-modified antibody fragments can provide a way to balance tumor penetration and accumulation with improved serum persistence...
April 10, 2018: Journal of Controlled Release: Official Journal of the Controlled Release Society
Shahrukh Husain, Vijay Kumar, Md Imtaiyaz Hassan
Tank binding kinase 1 (TBK-1) plays an important role in immunity, inflammation, autophagy, cell growth and proliferation. Nevertheless, a key molecular and structural detail of TBK-1 phosphorylation and activation has been largely unknown. Here we investigated the energy landscape of phosphorylated (active) and unphosphorylated (inactive) forms of human TBK-1 to characterize the interplay between phosphorylation and local frustration. By employing the algorithm equipped with energy function and implemented in Frustratometer web-server (http://www...
April 10, 2018: Journal of Theoretical Biology
Melissa Drappier, Babal Kant Jha, Sasha Stone, Ruth Elliott, Rong Zhang, Didier Vertommen, Susan R Weiss, Robert H Silverman, Thomas Michiels
The OAS/RNase L pathway is one of the best-characterized effector pathways of the IFN antiviral response. It inhibits the replication of many viruses and ultimately promotes apoptosis of infected cells, contributing to the control of virus spread. However, viruses have evolved a range of escape strategies that act against different steps in the pathway. Here we unraveled a novel escape strategy involving Theiler's murine encephalomyelitis virus (TMEV) L* protein. Previously we found that L* was the first viral protein binding directly RNase L...
April 13, 2018: PLoS Pathogens
Rong Chen, Shin-Ho Chung
Molecular dynamics simulations are employed to determine the inhibitory mechanisms of three drugs, 5-(4- phenoxybutoxy)psoralen (PAP-1), vernakalant and flecainide, on the voltage-gated K+ channel Kv1.5, a target for the treatment of cardiac arrhythmia. At neutral pH, PAP-1 is neutral, whereas the other two molecules carry one positive charge. We show that PAP-1 forms stable dimers in water, primarily through the hydrophobic interactions between aromatic rings. All three molecules bind to the cavity between the Ile508 and Val512 residues from the four subunits of the channel...
April 13, 2018: Biochemistry
C E Alvarez, F Trajtenberg, N Larrieux, M Saigo, A Golic, C S Andreo, S A Hogenhout, M A Mussi, M F Drincovich, A Buschiazzo
Phytoplasmas are wall-less phytopathogenic bacteria that produce devastating effects in a wide variety of plants. Reductive evolution has shaped their genome, with the loss of many genes, limiting their metabolic capacities. Owing to the high concentration of C4 compounds in plants, and the presence of malic enzyme (ME) in all phytoplasma genomes so far sequenced, the oxidative decarboxylation of L-malate might represent an adaptation to generate energy. Aster yellows witches'-broom (Candidatus Phytoplasma) ME (AYWB-ME) is one of the smallest of all characterized MEs, yet retains full enzymatic activity...
April 1, 2018: Acta Crystallographica. Section D, Structural Biology
Maxim Igaev, Helmut Grubmüller
Microtubules (MTs) are key components of the cytoskeleton and play a central role in cell division and development. MT assembly is known to be associated with a structural change in αβ-tubulin dimers from kinked to straight conformations. How GTP binding renders individual dimers polymerization-competent, however, is still unclear. Here, we have characterized the conformational dynamics and energetics of unassembled tubulin using atomistic molecular dynamics and free energy calculations. Contradictory to existing allosteric and lattice models, we find that GTP-tubulin favors a broad range of almost isoenergetic curvatures, whereas GDP-tubulin has a much lower bending flexibility...
April 13, 2018: ELife
Pelayo Camps, David Lozano, Carla Barbaraci, Merce Font-Bardia, F Javier Luque, Carolina Estarellas
Octacyclo[,10.03,7.04,9.08,19.011,16.013,17]nonadeca-5,8,14-triene (27) a hindered pyramidalized alkene, has been generated from a diiodide precursor. Contrary to the usual behavior of known pyramidalized alkenes, no Diels-Alder adducts were obtained from the present alkene when it was generated by different standard procedures in the presence of different dienes. However, products derived from the reduction, t-BuLi addition, condensation with the solvent, or dimerization, were isolated from these reactions, depending on the conditions used to generate it...
April 13, 2018: Journal of Organic Chemistry
Stephanie A Beach, Linda H Doerrer
As the scale of microelectronic circuit devices approaches the atomic limit, the study of molecular-based wires and magnets has become more prevalent. Compounds with quasi-1D geometries have been investigated for their electronic conductivity and magnetic properties with potential use as nanoscale circuit components and information storage devices. To increase the number of compositionally tailored molecular systems available to study, we have taken a building-block, bottom-up approach to the development of improved electronic structure and magnetic properties of quasi-1D arrays...
April 13, 2018: Accounts of Chemical Research
Anna I Sulatskaya, Georgy N Rychkov, Maksim I Sulatsky, Natalia P Rodina, Irina M Kuznetsova, Konstantin K Turoverov
The aim of the present work was investigation of the fluorescent dye thioflavin T (ThT) binding to acetylcholinesterase (AChE). ThT is an effectively used test for the protease activity, as well as a probe for amyloid fibril formation. Despite the extended and active investigation of the ThT - AChE binding, there is still no common view on the stoichiometry of this interaction. In particular, there is a hypothesis explaining the spectral properties of the bound to AChE dye and high quantum yield of its fluorescence by formation of dimers or excimers of ThT...
April 13, 2018: ACS Chemical Neuroscience
Yulin Tian, Qing Lin
Three γ-heteroatom-substituted N-methylpyrroletetrazole-lysines (mPyTXKs) were synthesized and subsequently incorporated into proteins site-specifically via genetic code expansion. The γ-seleno-substituted derivative, mPyTSeK, showed excellent incorporation efficiency in Escherichia coli and allowed site-selective photo-cross-linking of the GST dimer. Furthermore, the mPyTSeK-cross-linked GST dimer can be cleaved under mild oxidative conditions. The incorporation of mPyTXKs into proteins in mammalian cells was also demonstrated...
April 13, 2018: Chemical Communications: Chem Comm
Ezequiel Iván Juritz, Juan Pablo Bascur, Daniel Eduardo Almonacid, Fernando Danilo González-Nilo
BACKGROUND: Isocitrate dehydrogenase 1 (IDH1) is a dimeric enzyme responsible for supplying the cell's nicotinamide adenine dinucleotide phosphate (NADPH) reserves via dehydrogenation of isocitrate (ICT) and reduction of NADP+. Mutations in position R132 trigger cancer by enabling IDH1 to produce D-2-hydroxyglutarate (2-HG) and reduce inhibition by ICT. Mutant IDH1 can be found as a homodimer or a heterodimer. OBJECTIVE: We propose a novel strategy to inhibit IDH1 R132 variants as a means not to decrease the concentration of 2-HG but to provoke a cytotoxic effect, as the cell malignancy at this point no longer depends on 2-HG...
April 12, 2018: Molecular Diagnosis & Therapy
Javier E Moreno, Guillermo Moreno-Piovano, Raquel L Chan
The bHLH family is composed by canonical and non-canonical transcription factors (TFs) that differ in the presence or absence of their DNA-binding domain, respectively. Since both types of bHLH proteins are able to dimerize, their relative abundance impacts their biological activity. Among this TF family BEE and IBH are canonical and non-canonical bHLHs, respectively and previous reports indicated that BEE2 and IBH1 dimerize. Wondering whether BEE TFs participate in the abiotic stress response and how the dimerization with IBH1 could regulate their role in Arabidopsis, double bee1/bee2 and triple bee1/bee2/bee3 mutants were tested under salinity and drought stresses...
June 2018: Plant Science: An International Journal of Experimental Plant Biology
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