Biochemical characterization of an in-house Coccidioides antigen: perspectives for the immunodiagnosis of coccidioidomycosis

The objective of this study was to evaluate the reactivity of an in-house antigen, extracted from a strain of C. posadasii isolated in northeastern Brazil, by radial immunodiffusion and Western blotting, as well as to establish its biochemical characterization. The protein antigen was initially extracted with the use of solid ammonium sulfate and characterized by 1-D electrophoresis. Subsequently, it was tested by means of double radial immunodiffusion and Western blotting. A positive reaction was observed against the antigen by both immunodiagnostic techniques tested on sera from patients suffering from coccidioidomycosis. Besides this, two immunoreactive protein bands were observed and were revealed to be a β-glucosidase and a glutamine synthetase after sequencing of the respective N-terminal regions. Our in-house Coccidioides antigen can be promising as a quick and low-cost diagnostic tool without the risk of direct manipulation of the microorganism.     

Optimized measurement temperature gives access to the solution structure of a 49 kDa homohexameric β-propeller

Ph1500 is a homohexameric, two-domain protein of unknown function from the hyperthermophilic archaeon Pyrococcus horikoshii. The C-terminal hexamerization domain (Ph1500C) is of particular interest, as it lacks sequence homology to proteins of known structure. However, it resisted crystallization for X-ray analysis, and proteins of this size (49 kDa) present a considerable challenge to NMR structure determination in solution. We solved the high-resolution structure of Ph1500C, exploiting the hyperthermophilic nature of the protein to minimize unfavorable relaxation properties by high-temperature measurement. Thus, the side chain assignment (97%) and structure determination became possible at full proton density. To our knowledge, Ph1500C is the largest protein for which this has been achieved. To minimize detrimental fast water exchange of amide protons at increased temperature, we employed a strategy where the temperature was optimized separately for backbone and side chain experiments.     

An Atypical Naturally Split Intein Engineered for Highly Efficient Protein Labeling

Protein trans‐splicing catalyzed by split inteins is a powerful technique for assembling a polypeptide backbone from two separate parts. However, split inteins with robust efficiencies and short fragments suitable for peptide synthesis are rare and have mostly been artificially created. The novel split intein AceL‐TerL was identified from metagenomic data and characterized. It represents the first naturally occurring, atypically split intein. The N‐terminal fragment of only 25 amino acids is the shortest natural intein fragment to date and was easily amenable to chemical synthesis with a fluorescent label. Optimal protein trans‐splicing activity was observed at low temperatures. Further improved mutants were selected by directed protein evolution. The engineered intein variants with up to 50‐fold increased rates showed unprecedented efficiency in chemically labeling of a diverse set of proteins. These inteins should prove valuable tools for protein semi‐synthesis and other intein‐related biotechnological applications.     

Mathematical research data

The generation and processing of research data for modeling and simulation is an important part of the scientific work at WIAS. Together with scientific software it plays an essential role for the transfer of mathematical research results to industry or to other scientific disciplines. In the face of the emerging digital science agenda research data is now more and more recognized as an essential foundation for scientific work. The importance of research data is also emphasized by the recent adoption of the “DFG Guidelines on the Handling of Research Data” by the Deutsche Forschungsgemeinschaft and the Open Data Pilot within the EU Horizon 2020 programme. In this contribution we present our definition of research data in mathematical modeling and simulation and requirements for its management. (© 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim)     

A comparison of the eigenvalues of the Dirac and Laplace operators on a two-dimensional torus

We compare the eigenvalues of the Dirac and Laplace operator on a two-dimensional torus with respect to the trivial spin structure. In particular, we compute their variation up to order 4 upon deformation of the flat metric, study the corresponding Hamiltonian and discuss several families of examples. Received: 10 December 1998     

Alltagsvorstellungen und chemische Erklärungskonzepte: “Die Teilchen saugen das Aroma aus dem Tee”

Much recent research in the educational field has been concerned with finding out the ideas which learners typically hold to explain phenomena and fundamental ideas in chemistry. Some of these conceptions do not correspond with the scientific view, for example the transfer of properties from the level of substances onto the level of particles. Many students fall back on their original conceptions outside the classroom. Examples of those daily‐life concepts and their development are presented here.     

Eigenvalue estimates for Dirac operators with parallel characteristic torsion

Assume that the compact Riemannian spin manifold (Mn,g) admits a G-structure with characteristic connection ∇ and parallel characteristic torsion (∇T=0), and consider the Dirac operator D1/3 corresponding to the torsion T/3. This operator plays an eminent role in the investigation of such manifolds and includes as special cases Kostant's “cubic Dirac operator” and the Dolbeault operator. In this article, we describe a general method of computation for lower bounds of the eigenvalues of D1/3 by a clever deformation of the spinorial connection. In order to get explicit bounds, each geometric structure needs to be investigated separately; we do this in full generality in dimension 4 and for Sasaki manifolds in dimension 5.