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121.
Genetically Encoding Photoswitchable Click Amino Acids in Escherichia coli and Mammalian Cells
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Dr. Christian Hoppmann Dr. Vanessa K. Lacey Dr. Gordon V. Louie Jing Wei Prof. Joseph P. Noel Prof. Lei Wang 《Angewandte Chemie (International ed. in English)》2014,53(15):3932-3936
The ability to reversibly control protein structure and function with light would offer high spatiotemporal resolution for investigating biological processes. To confer photoresponsiveness on general proteins, we genetically incorporated a set of photoswitchable click amino acids (PSCaas), which contain both a reversible photoswitch and an additional click functional group for further modifications. Orthogonal tRNA‐synthetases were evolved to genetically encode PSCaas bearing azobenzene with an alkene, keto, or benzyl chloride group in E. coli and in mammalian cells. After incorporation into calmodulin, the benzyl chloride PSCaa spontaneously generated a covalent protein bridge by reacting with a nearby cysteine residue through proximity‐enabled bioreactivity. The resultant azobenzene bridge isomerized in response to light, thereby changing the conformation of calmodulin. These genetically encodable PSCaas will prove valuable for engineering photoswitchable bridges into proteins for reversible optogenetic regulation. 相似文献
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Dr. Moustafa Malki Dr. Santiago Casado Dr. María Francisca López Dr. Renaud Caillard Dr. F. Javier Palomares Dr. José A. Martín Gago Dr. Cristina Vaz‐Domínguez Dr. Angel Cuesta Prof. Ricardo Amils Prof. Víctor M. Fernández Dr. Marisela Velez Dr. Antonio L. De Lacey Dr. David Olea 《Chemphyschem》2013,14(6):1237-1244
The biofilm formation of a strain of the extremophile bacterium Acidiphilium sp., capable of donating electrons directly to electrodes, was studied by different surface characterization techniques. We develop a method that allows the simultaneous study of bacterial biofilms by means of fluorescence microscopy and atomic force microscopy (AFM), in which transparent graphitic flakes deposited on a glass substrate are used as a support for the biofilm. The majority of the cells present on the surface were viable, and the growth of the biofilms over time showed a critical increase of the extracellular polymeric substances (EPS) as well as the formation of nanosized particles inside the biofilm. Also, the presence of Fe in Acidiphilium biofilms was determined by X‐ray photoelectron spectroscopy (XPS), whereas surface‐enhanced infrared absorption spectroscopy indicated the presence of redox‐active proteins. 相似文献
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Two series of liquid crystalline cyclic siloxane tetramers, one containing the 2-methylbutyl chiral group and the other the 1-methylheptyl chiral group, were prepared to investigate, in a systematic manner, the role of molecular structure of (a) the spacer group, (b) the mesogenic side chain and (c) the chiral end group, on the liquid crystalline behaviour of these novel tetramers. The results from this systematic structure/property correlation study clearly showed the effect of the structure of both the chiral end group and the mesogenic side chain core on the thermal properties and temperature ranges of the SmC* phase (ferroelectric) exhibited by these novel materials. By the appropriate choice of spacer group, mesogenic side chain and chiral end group, a number of cyclic siloxane tetramers exhibiting wide SmC* ranges (ferroelectricity) around room temperature were synthesized. 相似文献
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Niu Y Padhee S Wu H Bai G Harrington L Burda WN Shaw LN Cao C Cai J 《Chemical communications (Cambridge, England)》2011,47(44):12197-12199
We report the identification of a new class of antimicrobial peptidomimetics-γ-AApeptides with potent and broad-spectrum activity, including clinically-relevant strains that are unresponsive to most antibiotics. They are also not prone to select for drug-resistance. 相似文献
128.
Liebgott PP de Lacey AL Burlat B Cournac L Richaud P Brugna M Fernandez VM Guigliarelli B Rousset M Léger C Dementin S 《Journal of the American Chemical Society》2011,133(4):986-997
Hydrogenases are efficient biological catalysts of H(2) oxidation and production. Most of them are inhibited by O(2), and a prerequisite for their use in biotechnological applications under air is to improve their oxygen tolerance. We have previously shown that exchanging the residue at position 74 in the large subunit of the oxygen-sensitive [NiFe] hydrogenase from Desulfovibrio fructosovorans could impact the reaction of the enzyme with O(2) (Dementin, S.; J. Am. Chem. Soc. 2009, 131, 10156-10164; Liebgott, P. P.; Nat. Chem. Biol. 2010, 6, 63-70). This residue, a valine in the wild-type enzyme, located at the bottleneck of the gas channel near the active site, has here been exchanged with a cysteine. A thorough characterization using a combination of kinetic, spectroscopic (EPR, FTIR), and electrochemical studies demonstrates that the V74C mutant has features of the naturally occurring oxygen-tolerant membrane-bound hydrogenases (MBH). The mutant is functional during several minutes under O(2), has impaired H(2)-production activity, and has a weaker affinity for CO than the WT. Upon exposure to O(2), it is converted into the more easily reactivatable inactive form, Ni-B, and this inactive state reactivates about 20 times faster than in the WT enzyme. Control experiments carried out with the V74S and V74N mutants indicate that protonation of the position 74 residue is not the reason the mutants reactivate faster than the WT enzyme. The electrochemical behavior of the V74C mutant toward O(2) is intermediate between that of the WT enzyme from D. fructosovorans and the oxygen-tolerant MBH from Aquifex aeolicus. 相似文献
129.
Han S Caspers N Zaniewski RP Lacey BM Tomaras AP Feng X Geoghegan KF Shanmugasundaram V 《Journal of the American Chemical Society》2011,133(50):20536-20545
Multi-drug-resistant forms of the Gram-negative pathogen Acinetobacter baumannii are an emerging threat to human health and further complicate the general problem of treating serious bacterial infections. Meeting this challenge requires an improved understanding of the relationships between the structures of major therapeutic targets in this organism and the activity levels exhibited against it by different antibiotics. Here we report the first crystal structures of A. baumannii penicillin-binding proteins (PBPs) covalently inactivated by four β-lactam antibiotics. We also relate the results to kinetic, biophysical, and computational data. The structure of the class A protein PBP1a was solved in apo form and for its covalent conjugates with benzyl penicillin, imipenem, aztreonam, and the siderophore-conjugated monocarbam MC-1. It included a novel domain genetically spliced into a surface loop of the transpeptidase domain that contains three conserved loops. Also reported here is the first high-resolution structure of the A. baumannii class B enzyme PBP3 in apo form. Comparison of this structure with that of MC-1-derivatized PBP3 of Pseudomonas aeruginosa identified differences between these orthologous proteins in A. baumannii and P. aeruginosa. Thermodynamic analyses indicated that desolvation effects in the PBP3 ligand-binding sites contributed significantly to the thermal stability of the enzyme-antibiotic covalent complexes. Across a significant range of values, they correlated well with results from studies of inactivation kinetics and the protein structures. The structural, biophysical, and computational data help rationalize differences in the functional performance of antibiotics against different protein targets and can be used to guide the design of future agents. 相似文献
130.
We prove a vector-valued version of Carleson’s theorem: let $Y=[X,H]_\theta $ be a complex interpolation space between an unconditionality of martingale differences (UMD) space $X$ and a Hilbert space $H$ . For $p\in (1,\infty )$ and $f\in L^p(\mathbb T ;Y)$ , the partial sums of the Fourier series of $f$ converge to $f$ pointwise almost everywhere. Apparently, all known examples of UMD spaces are of this intermediate form $Y=[X,H]_\theta $ . In particular, we answer affirmatively a question of Rubio de Francia on the pointwise convergence of Fourier series of Schatten class valued functions. 相似文献