Technische Universität Berlin - Faculty III - Institute of Biotechnology / Chair of Bioanalytics
Research Assistant - 0.67 working time - salary grade E13 TV-L Berliner Hochschulen
under the reserve that funds are granted
The focus of the DFG-funded project "Site Specific Crosslinking with KLK Proteases from the Prostate" (BU 1404 12-1) is on the development of methods for the detection and identification of short-lived protein-ligand complexes difficult to access with classical chemical modifications. In particular, non-canonical amino acid (ncAAs) based click chemistry will be used to study protein-ligand interactions in human kallikrein-related peptidases (KLKs) from the prostate (especially KLKs 2, 3, 4). The main task is to develop a site-specific / statistical cross-linking of proteins with ncAAs, which should generally be applicable to protein complexes with different ligands. The specific goal is to determine enzymatic substrate specificity (in the concrete case, "tertiary specificity" of the KLK proteases) useful for elucidating biological and disease-related processes, e.g. contribute to prostate cancer and allow the design of new drugs. Parallel to work with recombinant KLK protease and chemical synthesis of desired ncAAs, the chemical synthesis of suitable ligands will be performed as well. Basic analytical (amino acid analysis, various mass spectrometric methods, etc.) and biophysical methods (NMR, CD spectroscopy, fluorescence and UV absorbance, etc.) find a very strong application throughout the project. Biosynthetic work includes enzyme evolution for novel substrates, design of orthogonal pairs, establishment of orthogonal pairs in bacterial and eukaryotic cells, and subsequent optimization of expression and measurement conditions. The incorporation of the ncAAs will be carried out by means of various orthogonal pairs or in combination with the selection pressure incorporation (SPI) method (statistical and site-directed labelling).
Successfully completed university degree (Master, Diplom or equivalent) preferably in chemistry, biotechnology, chemical biology or a related field; a very good grade average desired. Candidates who are about to graduate are also invited to apply. The generation of suitable expression systems requires extensive knowledge in the field of molecular biology (cloning methods, cloning software such as Geneious / Clone Manager / APE, expression vectors and strains). Protein and peptide purification require very good knowledge of the common chromatographic methods (see above) and the handling of the Äkta systems available at the institute. For the interpretation of u. a. MS spectra, activity and fluorescence assays require good computer skills in the area of data analysis (Excel, Origin / SigmaPlot) and protein structure analysis (Pymol / Chimera / Swiss PDB viewer or similar). Experience in the analysis of proteins by means of crosslinking mass spectrometry as well as in chemical synthesis of amino acids including peptide synthesis is especially advantageous.
The capacity to communicate with various specialists in the fields of synthetic chemistry, molecular biology and bio-medicine is a prerequisite. Openness for interdisciplinary research projects in the field of natural sciences and high motivation for independent scientific work ideally with (if possible) proven success through publications; very good communication and team skills and excellent command of spoken and written English.
How to apply:
Please send your written application with the reference number and the usual documents to Technische Universität Berlin - Der Präsident - Fakultät III, Institut für Biotechnologie, FG Bioanalytik, Prof. Dr. Rappsilber, Sekr. TIB 4/4-3, Gustav-Meyer-Allee 25, 13355 Berlin or by e-mail to email@example.com.
To ensure equal opportunities between women and men, applications by women with the required qualifications are explicitly desired. Qualified individuals with disabilities will be favored. The TU Berlin values the diversity of its members and is committed to the goals of equal opportunities.
Please send copies only. Original documents will not be returned.