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181.
Dr. Ivana Nikić Dr. Tilman Plass Dr. Oliver Schraidt Dr. Jędrzej Szymański Dr. John A. G. Briggs Priv.‐Doz. Dr. Carsten Schultz Dr. Edward A. Lemke 《Angewandte Chemie (International ed. in English)》2014,53(8):2245-2249
The growing demands of advanced fluorescence and super‐resolution microscopy benefit from the development of small and highly photostable fluorescent probes. Techniques developed to expand the genetic code permit the residue‐specific encoding of unnatural amino acids (UAAs) armed with novel clickable chemical handles into proteins in living cells. Here we present the design of new UAAs bearing strained alkene side chains that have improved biocompatibility and stability for the attachment of tetrazine‐functionalized organic dyes by the inverse‐electron‐demand Diels–Alder cycloaddition (SPIEDAC). Furthermore, we fine‐tuned the SPIEDAC click reaction to obtain an orthogonal variant for rapid protein labeling which we termed selectivity enhanced (se) SPIEDAC. seSPIEDAC and SPIEDAC were combined for the rapid labeling of live mammalian cells with two different fluorescent probes. We demonstrate the strength of our method by visualizing insulin receptors (IRs) and virus‐like particles (VLPs) with dual‐color super‐resolution microscopy. 相似文献
182.
Light‐Induced Conformational Changes in the Plant Cryptochrome Photolyase Homology Region Resolved by Selective Isotope Labeling and Infrared Spectroscopy 下载免费PDF全文
Constanze Sommer Marina S. Dietz Thomas Patschkowski Tilo Mathes Tilman Kottke 《Photochemistry and photobiology》2017,93(3):881-887
Plant cryptochromes are photoreceptors that regulate flowering, circadian rhythm and photomorphogenesis in response to blue and UV‐A light. It has been demonstrated that the oxidized flavin cofactor is photoreduced to the neutral radical state via separate electron and proton transfer. Conformational changes have been found in the C‐terminal extension, but few studies have addressed the changes in secondary structure in the sensory photolyase homology region (PHR). Here, we investigated the PHR of the plant cryptochrome from the green alga Chlamydomonas reinhardtii by light‐induced infrared difference spectroscopy in combination with global 13C and 15N isotope labeling. Assignment of the signals is achieved by establishing a labeling strategy for cryptochromes that preserves the flavin at natural abundance. We demonstrate by UV/vis spectroscopy that the integrity of the sample is maintained and by mass spectrometry that the global labeling was highly efficient. As a result, difference bands are resolved at full intensity that at natural abundance are compensated by the overlap of flavin and protein signals. These bands are assigned to prominent conformational changes in the PHR by blue light illumination. We postulate that not only the partial unfolding of the C‐terminal extension but also changes in the PHR may mediate signaling events. 相似文献
183.
Joseph A. Piccirilli Tilman Krauch Lawrence J. MacPherson Steven A. Benner 《Helvetica chimica acta》1991,74(2):397-406
A route for synthesizing C-nucleosides with 2,6-substituted pyridines as heterocyclic aglycones is described. Condensation of appropriately substituted lithiated pyridines with ribono-1,4-lactone derivatives yields hemiacetal 4a – g (Table 1), which can be reduced by Et3SiH and BF3·Et2O to the corresponding C-nucleoside (see Scheme 1 for 4d → β-D - 5 ). Conditions are presented that optimize the amount of the 2,6-dichloropyridine-derived β-D -anomer β-D - 5 formed (Table 3). Aminolysis of β-D - 5 yields the diaminonucleoside 14 (Scheme 3). 相似文献
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185.
In order to facilitate model reduction by balanced truncation, we introduce state space transformations that can be used to construct an ℓ2-balanced realization of a regular, linear input-ouput map with nuclear Hankel-operator directly from the system generators of an arbitrary, given realization. These balancing transformations are based on factors of the Gramians and, for infinite-dimensional systems, they are usually unbounded operators. Subsequently the ℓ2-balanced realization can be truncated in a non-trivial way to obtain an approximating, finite-dimensional model. (© 2013 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim) 相似文献