Project 1: Carbohydrate active enzymes in S-layer protein glycosylation
Several human pathogens contain glycoproteins as surface decoration. These compounds possess an enormous potential as lead structures for drug discovery and open a new arena of biomedical research.
This project focuses on the pathogenicity aspect of the surface (S-) layer glycosylation of the Gram-negative oral pathogen Tannerella forsythia. T. forsythia has been implicated in the development of periodontitis, which is one of the most common inflammatory diseases amongst the adult population. Thus, there is a great biomedical interest in understanding the host-bacteria cross-talk that forms the basis of health, disease, and healing. While there is already clear evidence that the S layer of T. forsythia is a virulence factor, we hypothesize that the glycans attached to the S-layer are keys to the virulence of this bacterium. To initiate this direction of research, the enzymes involved in S-layer glycosylation of T. forsythia shall be characterized.
-Identification of carbohydrate-active candidate enzymes based on the ORFs annotated in the Tannerella genome
-Cloning, over-expression and purification of the enzymes
-Set-up of functional enzyme assays and kinetics measurements
-Structure analysis of the enzymes (e.g., circular dichroism, Fourier-transform IR spectroscopy)
-3D crystallization of selected enzymes
Project 2: Self-assembly of S-layer glycoproteins for applications in nanobiotechnology
Self-assembly of molecules into crystalline arrays and organizing matter at the nano-scale level are new and rapidly growing transdisciplinary scientific and engineering fields. Bacterial surface (S-) layers are periodic structures formed by self-assembly. The current project focuses on S-layers that are naturally glycosylated and, thus, provide a unique setting to exploit glycan-mediated, biological phenomena such as recognition, signalling, trafficking, biological half-life time, or adhesion. The self-assembly process of S-layer glycoproteins shall be characterized in vivo, where protein self-assembly has to be coordinated with the glycosylation process, and in vitro, as a basis for applications of glycan-functionalized S-layer proteins in nanobiotechnology.
-Understanding of the in vitro self-assembly process (scanning probe microscopy, surface plasmon resonance, quartz crystal microbalance measurements with dissipation monitoring)
-Study of intermolecular interactions within S-layer lattices (e.g., force spectroscopy and steered molecular dynamic simulations)
-In vivo S-layer glycoprotein folding and assembly
-Interactions studies of S-layer glycoprotein lattices with selected biomolecules (e.g., QCM-D, SPR.)
The candidate must hold a MSc degree and should have a sound background in molecular biology, microbiology, and biochemistry. Practical experience in the field of enzymology and glycobiology is of advantage but not mandatory. The highly motivated candidate should be an excellent team player with the ability to work independently in an interdisciplinary environment.
For detailed information on research projects, application guidelines and download of forms please visit: http://biotop.boku.ac.at
Application deadline: September 30th, 2010
Anfangsdatum: 1. März 2011
geschätzte Dauer: 3 years