- Journal article
Authors : Mazurek, Łukasz; Ghilarov, Dmitry; Michalczyk, Elizabeth (2020) - Activities of two such proteins, QnrB1 and AlbG in vitro. Each of them provided specific protection against its cognate toxin (fluoroquinolone or albicidin), which strictly required ATP hydrolysis by gyrase. Through a combination of fluorescence anisotropy, pull-downs and photocrosslinking we show that QnrB1 binds to the GyrB protein. We further probed the QnrB1 binding site using site-specific incorporation of a photoreactive amino acid and mapped strong and...
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- Periodicals (Báo – Tạp chí)
Authors : Moroz-Omori, Elena V; Hoàng, Đan Chi (2020) - The methylase METTL3 is the writer enzyme of the N6-methyladenosine (m6A) modification of RNA. Using a structure-based drug discovery approach, we identified a METTL3 inhibitor (UZH1a) with potency in a biochemical assay of 280 nM, while its enantiomer UZH1b is 100 times less active. The crystal structure of the complex of METTL3 with UZH1a illustrates the interactions that make it selective against protein methyltransferases. We observed a dose-dependen...
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- Periodicals (Báo – Tạp chí)
Authors : Puri, Neha (2020) - To dissect how specific ATPase elements of E. coli DnaC, an archetypal loader for the bacterial DnaB helicase, play distinct roles in helicase loading and the activation of DNA unwinding. We identify a new element, the arginine-coupler, which regulates the switch-like behavior of DnaC to prevent futile ATPase cycling and maintains loader responsiveness to replication restart systems. Our data help explain how the ATPase cycle of a AAA+-family helica...
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- Periodicals (Báo – Tạp chí)
Authors : Ngo, Tuan-Dung (2020) - A protein-protein interaction ELISA revealed the interaction of PscN with a wide range of exported T3SS proteins including the needle, translocator, gate-keeper and effector. These interactions were further confirmed by Microscale Thermophoresis that also indicated a preferential interaction of PscN with secreted proteins or protein-chaperone complex rather than with chaperones alone, in line with the release of the chaperones in the bacterial cytoplasm after the ...
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- Periodicals (Báo – Tạp chí)
Authors : Brittain, Tyler J. (2020) - Results on expression and biophysical characterization of GSU0105, a novel 3-heme cytochrome, important for Fe(III) respiration in G. sulfurreducens. We successfully cloned the gene and achieved ~3 mg/L of culture GSU0105 expression in E.coli. Despite a similar size (71 amino acids) and the same number of c-type hemes to the members of the cytochrome (cyt) c7 family, multiple sequence alignment suggests that GSU0105 does not belong to the cyt&#...
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- Periodicals (Báo – Tạp chí)
Authors : Syson, Karl (2020) - To reconstruct and examine the temperature-sensitive mutant and characterise the mutated GlgE enzyme. Results- The mutant strain accumulated the substrate for GlgE, -maltose-1-phosphate, at the non-permissive temperature. The glycogen assay used in the original study was shown to give a false positive result with -maltose-1-phosphate. The accumulation of -maltose-1-phosphate was due to the lowering of the kcat of GlgE as well as a loss of stability 42 ...
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- Journal article
Authors : Menon, Nikhil G.; Goyal, Ruchi; Lema, Carolina (2020) - Improve signs and symptoms of DED in humans. The objectives of this study were to characterize endogenous PRG4 expression by telomerase-immortalized human corneal epithelial (hTCEpi) cells, examine how exogenous recombinant human PRG4 (rhPRG4) modulates cytokine and chemokine secretion in response to TNFα and IL-1β, explore rhPRG4 as a potential substrate and/or inhibitor of MMP-9, and to understand how experimental dry (EDE) in mice affects PRG4 expression.<...
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- Periodicals (Báo – Tạp chí)
Authors : Jooss, Kevin (2020) - Provide standard operating procedures for acquiring high quality data using CZE in native mode coupled online to various Orbitrap mass spectrometers via a commercial sheathless interface, covering a wide range of analytes from 30 – 800 kDa. Using a standard protein mix, the influence of various CZE method parameters were evaluated, such as BGE/conductive liquid composition and separation voltage. Additionally, a universal approach for the optimization of...
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- Journal article
Authors : Luong, Jennie A.; Vater, Ashley; Siegel, Justin B. (2020) - The Michaelis-Menten constants (kcat, KM, and kcat/KM) and thermal stability (TM) of five new mutations of β-glucosidase B from Paenibacillus polymyxa (BglB) are characterized. Foldit software was used to create molecular models of the mutants, for which synthetic genes were constructed and the corresponding proteins produced and purified from E. coli. It was found that mutations that disrupted pre-existing hydrogen bonds near the active site had reduced...
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- Journal article
Authors : Malcolm, Tess R.; Belousoff, Matthew J.; Venugopal, Hariprasad (2020) - The active site metal ions essential for catalytic activity have a secondary structural role mediating the formation of active hexamers. We found that PfA-M17 and Pv-M17 exist in a metal-dependent dynamic equilibrium between active hexameric species and smaller inactive species, that can be controlled by manipulating the identity and concentration of metal ions available. Mutation of residues involved in metal ion binding impaired catalytic activity and the&#...
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