Kiri Chr. W. F. Walchile, Göttingen

http://tartu.ester.ee/record=b1881198~S1*es

Structural analysis of the adenovirus type 2 E3/19K protein using mutagenesis and a panel of conformation-sensitive monoclonal antibodies

The E3/19K protein of human adenovirus type 2 (Ad2) was the first viral protein shown to interfere with antigen presentation. This 25 kDa transmembrane glycoprotein binds to major histocompatibility complex (MHC) class I molecules in the endoplasmic reticulum (ER), thereby preventing transport of newly synthesized peptide–MHC complexes to the cell surface and consequently T cell recognition. Recent data suggest that E3/19K also sequesters MHC class I like ligands intracellularly to suppress natural killer (NK) cell recognition. While the mechanism of ER retention is well understood, the structure of E3/19K remains elusive. To further dissect the structural and antigenic topography of E3/19K we carried out site-directed mutagenesis and raised monoclonal antibodies (mAbs) against a recombinant version of Ad2 E3/19K comprising the lumenal domain followed by a C-terminal histidine tag. Using peptide scanning, the epitopes of three mAbs were mapped to different regions of the lumenal domain, comprising amino acids 3–13, 15–21 and 41–45, respectively. Interestingly, mAb 3F4 reacted only weakly with wild-type E3/19K, but showed drastically increased binding to mutant E3/19K molecules, e.g. those with disrupted disulfide bonds, suggesting that 3F4 can sense unfolding of the protein. MAb 10A2 binds to an epitope apparently buried within E3/19K while that of 3A9 is exposed. Secondary structure prediction suggests that the lumenal domain contains six β-strands and an α-helix adjacent to the transmembrane domain. Interestingly, all mAbs bind to non-structured loops. Using a large panel of E3/19K mutants the structural alterations of the mutations were determined. With this knowledge the panel of mAbs will be valuable tools to further dissect structure/function relationships of E3/19K regarding down regulation of MHC class I and MHC class I like molecules and its effect on both T cell and NK cell recognition

Fabrication of a single sub-micron pore spanning a single crystal (100) diamond membrane and impact on particle translocation

The fabrication of sub-micron pores in single crystal diamond membranes, which span the entirety of the membrane, is described for the first time, and the translocation properties of polymeric particles through the pore investigated. The pores are produced using a combination of laser micromachining to form the membrane and electron beam induced etching to form the pore. Single crystal diamond as the membrane material, has the advantages of chemical stability and durability, does not hydrate and swell, has outstanding electrical properties that facilitate fast, low noise current-time measurements and is optically transparent for combined optical-conductance sensing. The resulting pores are characterized individually using both conductance measurements, employing a microcapillary electrochemical setup, and electron microscopy. Proof-of-concept experiments to sense charged polystyrene particles as they are electrophoretically driven through a single diamond pore are performed, and the impact of this new pore material on particle translocation is explored. These findings reveal the potential of diamond as a platform for pore-based sensing technologies and pave the way for the fabrication of single nanopores which span the entirety of a diamond membrane

Die Letzte Pflicht, Wolten, Bey dem unvermutheten, doch seeligen Absterben, Des Hoch-Edlen, Groß-Achtbahren und Wohlweisen Herrn, Herrn Johann Friderich Oehlers, Eines Hoch-Edlen Magistrats zu Wernigerode treu-gewesenen und wohl-meritirten Cämmerers, Auch ... Kauf- u. Handels-Manns, Als Derselbe Am 13. Maji 1739. durch einen seeligen Tod der Zeitlichkeit entnommen, Und Dessen entseelter Cörper am 18. ejusd. zu seiner Ruhe-Satt [!] gebracht wurde, wehmüthigst bezeugen Dessen über diesen Todes-Fall schmertzlich betrübte Jnnen Benahmte

Autopsie nach Ex. der ULB Sachsen-AnhaltVorlageform des Erscheinungsvermerks: Wernigerode, Druckts Michael Anton Struck, Hoch-Gräfl. Stolbergl. Hof-Buchdrucker

Bey dem Absterben Der Hochedlen und Tugendreichen Frauen Sabina Catharina Frickin, gebohrnen Bornemannin

Wollten ihre Wehmuth bezeigen zwey Vettern Johann Christian von Windheim; Justus von WindheimAutopsie nach Ex. der ULB Sachsen-AnhaltVorlageform des Erscheinungsvermerks: Halle im Magdeburgischen 1740. gedruckt bey Johann Justinus Gebauer, Universitätsbuchdrucker

Auf das frühzeitige Absterben des Weiland Hoch-Edlen und Großachtbaren Herrn, Herrn Johann Friederich Oehler, Wohlbestalten Raths-Cämmerers der Stadt Wernigerode und fürnehmen Kauff- und Handelsmann daselbst, welcher den 13. Mai dieses Jahrs 1739. nach einer kurtzen Kranckheit, ... eingeschlaffen, und ... den 19. ejusd. ... beygesetzet worden; Haben ihre traurige Gedancken richten wollen ... im Leben liebgewesene Vettern Christian Ernst von Windheim; Rudolph August von Windheim

Autopsie nach Ex. der ULB Sachsen-AnhaltVorlageform des Erscheinungsvermerks: Braunschweig, gedruckt mit Keitelschen Schrifften

Den allzufrühzeitigen Verlust Ihrer innnigst geliebtesteb Frauen Mutter Frauen Dorotheen Augusten Margarethen von Windheim gebohrnen von Mosheim Welche den 27 Merz 1761 Abends um 10 Uhr das Zeitige mit dem Ewigen plötzlich verwechselte beweinen mit den heissesten Zähren der Wohlseligen beyde Kinder Johann August von Windheim; Henriette Auguste Dorothee von Windheim

Autopsie nach Ex. der ULB Sachsen-AnhaltVorlageform des Erscheinungsvermerks: Erlangen, gedruckt mit Wolfgang Waltherischen Schriften