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Proteolysis of SNAP‐25 Isoforms by Botulinum Neurotoxin Types A, C, and E
TLDR
It is demonstrated that cleavage of SNAP‐25 occurs between Arg198 and Ala199, depends on the presence of regions Asn93 to Glu145 and Ile156 to Met202, and requires about 1,000‐fold higher L chain concentrations in comparison with BoNT/A and BoNT /E. Expand
Light-induced flickering of DsRed provides evidence for distinct and interconvertible fluorescent states.
TLDR
Studying the emission dynamics of the recently cloned red fluorescent protein DsRed by fluorescence correlation spectroscopy (FCS), it can be verified that this protein exhibits strong light-dependent flickering similar to what is observed in several yellow-shifted mutants of GFP. Expand
Triple FRET: a tool for studying long-range molecular interactions.
An evolved aminoacyl-tRNA synthetase with atypical polysubstrate specificity.
TLDR
It is discovered that a p-cyanophenylalanine specific aminoacyl-tRNA synthetase (pCNF-RS) has high substrate permissivity for unnatural amino acids, while maintaining its ability to discriminate against the 20 canonical amino acids. Expand
Site-specific labeling of proteins with NMR-active unnatural amino acids
TLDR
A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions, and unique opportunities for site-specific labeling of proteins for NMR studies are discussed. Expand
Outwitting EF-Tu and the ribosome: translation with d-amino acids
TLDR
An in vitro translation system that enables single incorporation of d-aa into a polypeptide is introduced and reveals an unexpected plasticity of the ribosomal peptidyltransferase center and sheds new light on the mechanism of chiral discrimination during translation. Expand
Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
TLDR
FCS analyzes spontaneous fluctuations in the fluorescence emission of small molecular ensembles, thus providing information about a multitude of parameters, such as concentrations, molecular mobility and dynamics of fluorescently labeled molecules, which provides an attractive alternative for determining intracellular mobility parameters of very low quantities of fluorophores. Expand
A thermostable d-polymerase for mirror-image PCR
TLDR
It is believed that this d-polymerase provides a valuable tool to further investigate the mysteries of biological (homo)chirality and to pave the way for potential novel life forms running on a mirror-image genome. Expand
A New Embedded Process for Compartmentalized Cell‐Free Protein Expression and On‐line Detection in Microfluidic Devices
TLDR
This work demonstrates the efficient implementation of microstructured devices to generate water-in-oil emulsions, and at the same time perform in vitro expression of proteins inside the water droplets, using the red-shifted spectroscopy system. Expand
Triple-color coincidence analysis: one step further in following higher order molecular complex formation.
TLDR
It is shown that under certain experimental conditions, employing simultaneous two-photon excitation of three distinct dye species, their successful discrimination indeed becomes possible even on a single molecule level, and enables the direct observation of higher order molecular complex formation in the confocal volume. Expand
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