- Photolithographic Synthesis of Peptoids
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We describe a novel photolithographic approach to the synthesis of peptoids (oligo-N-substituted glycines). This strategy enables the construction of a spatially addressable peptoid microarray, thus providing a potentially powerful tool for the discovery
- Li, Shuwei,Bowerman, Dawn,Marthandan, Nishanth,Klyza, Stanley,Luebke, Kevin J.,Garner, Harold R.,Kodadek, Thomas
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- Genetically encoded photocontrol of protein localization in mammalian cells
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(Figure Presented) Precise photochemical control of protein function can be achieved through the site-specific introduction of caging groups. Chemical and enzymatic methods, including in vitro translation and chemical ligation, have been used to photocage proteins in vitro. These methods have been extended to allow the introduction of caged proteins into cells by permeabilization or microinjection, but cellular delivery remains challenging. Since lysine residues are key determinants for nuclear localization sequences, the target of key post-translational modifications (including ubiquitination, methylation, and acetylation), and key residues in many important enzyme active sites, we were interested in photocaging lysine to control protein localization, post-translational modification, and enzymatic activity. Photochemical control of these important functions mediated by lysine residues in proteins has not previously been demonstrated in living cells. Here we synthesized 1 and evolved a pyrrolysyl-tRNA synthetase/tRNA pair to genetically encode the incorporation of this amino acid in response to an amber codon in mammalian cells. To exemplify the utility of this amino acid, we caged the nuclear localization sequences (NLSs) of nucleoplasms and the tumor suppressor p53 in human cells, thus mislocalizing the proteins in the cytosol. We triggered protein nuclear import with a pulse of light, allowing us to directly quantify the kinetics of nuclear import. Copyright
- Gautier, Arnaud,Nguyen, Duy P.,Lusic, Hrvoje,An, Wenlin,Deiters, Alexander,Chin, Jason W.
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- Light Regulation of Enzyme Allostery through Photo-responsive Unnatural Amino Acids
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Imidazole glycerol phosphate synthase (ImGPS) is an allosteric bienzyme complex in which substrate binding to the synthase subunit HisF stimulates the glutaminase subunit HisH. To control this stimulation with light, we have incorporated the photo-respons
- Kneuttinger, Andrea C.,Straub, Kristina,Bittner, Philipp,Simeth, Nadja A.,Bruckmann, Astrid,Busch, Florian,Rajendran, Chitra,Hupfeld, Enrico,Wysocki, Vicki H.,Horinek, Dominik,K?nig, Burkhard,Merkl, Rainer,Sterner, Reinhard
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p. 1501 - 9,1514
(2019/11/20)
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- GENETICALLY ENCODED PHOTO CONTROL
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The invention relates to a caged lysine, wherein the caged lysine is according to Formula (I): or salts thereof. The invention further relates to polypeptides comprising a caged lysine, and to methods of making same. The invention further relates to tRNA synthetases capable of charging tRNA with caged lysine.
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Page/Page column 9
(2013/02/28)
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- The efficiency of light-directed synthesis of DNA arrays on glass substrates
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New methods based on photolithography and surface fluorescence were used to determine photodeprotection rates and stepwise yields for light-directed oligonucleotide synthesis using photolabile 5'-(((α-methyl-2- nitropiperonyl)-oxy)carbonyl)(MeNPOC)-2'-deoxynucleoside phosphoramidites on planar glass substrates. Under near-UV illumination (primarily 365 nm) from a mercury light source, the rate of photoremoval of the MeNPOC protecting group was found to be independent of both the nucleotide and length of the growing oligomer (t( 1/4 ) = 12 s at 27.5 mW/cm2). A moderate dependence on solvent polarity was observed, with photolysis proceeding most rapidly in the presence of nonpolar solvents or in the absence of solvent (e.g., t( 1/4 ) = 10 - 13 s at 27.5 mW/cm2). In solution, the photolysis rate was linearly dependent on light intensity over the range 5-50 mW/cm2. Average stepwise yields for the synthesis of dodecamer oligonucleotides were in the range of 92-94%, using monomers based on N6-(phenoxyacetyl)-2'-deoxyadenosine, N2- isobutyryl-2'-deoxyguanosine, N4-isobutyryl-2'-deoxycytidine, and thymidine. By comparison, an efficiency of 98%/step was obtained using a conventional 5'-dimethoxytrityl monomer with acid deprotection on the same support. The lower yields associated with the photochemical process appears to be due to incomplete recovery of free 5'-hydroxyl groups after photolysis on the support, although high yields of 5'-OH nucleosides (≤96%) are consistently observed when 5'-MeNPOC monomers are photolyzed in solution.
- McGall, Glenn H.,Barone, Anthony D.,Diggelmann, Martin,Fodor, Stephen P. A.,Gentalen, Erik,Ngo, Nam
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p. 5081 - 5090
(2007/10/03)
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