In the central project Z03 we have established a robust proteomics platform that allows us to combine specific purification approaches with quantitative proteomics in the coming funding period. Combining protein ligation techniques for immobilization of target proteins with 18O/16O-labelling will extend the interactome analysis in the coming funding period to primary cells and organotypic tissue cultures in several projects of the CRC. Detection of post-translational modifications such as palmitoylation or phosphorylation can be combined with the quantitative methods to yield compositional information on membrane scaffolds.
C. Barucker, A. Sommer, G. Beckmann, M. Eravci, A. Harmeier, C.G. Schipke, D. Brockschnieder, T. Dyrks, V. Althoff, P.E. Fraser, L.N. Hazrati, P.S. George-Hyslop, J.C. Breitner, O. Peters and G. Multhaup, J. Alzheimers Dis. 2015, 44, 613-24.
Kuropka, B., Royla, N., Freund, C., Krause, E. (2015) Sortase A mediated site-specific immobilization for identification of protein interactions in affinity purification-mass spectrometry experiments. Proteomics. 15, 1230-1234.
Stephanowitz, H., Lange, S., Lang, D., Freund, C. & Krause, E. An improved two-dimensional reversed phase-reversed phase LC-MS/MS approach for identification of peptide-protein interactions. J. Proteome Res. 11, 1175-83 (2012)
Eravci, M., Sommer, C.& Selbach, M. IPG strip-based peptide fractionation for shotgun proteomics. Methods Mol. Biol. 1156, 67-77 (2014)
Lange, S., Sylvester, M., Schümann, M., Freund, C. & Krause, E. Identification of phosphorylation-dependent interaction partners of the adapter protein ADAP using quantitative mass spectrometry: SILAC vs (18)O-labeling. J. Proteome Res. 9, 4113-22 (2010)
Lehmann, R., Meyer, J., Schuemann, M., Krause, E. & Freund, C. A novel S3S-TAP-tag for the isolation of T-cell interaction partners of adhesion and degranulation promoting adaptor protein. Proteomics 9, 5288-5295 (2009)
Kofler, M., Schümann, M., Merz, C., Kosslick, D., Schlundt, A., Tannert, A. Schaefer, M., Lührmann, R., Krause, E. and Freund, C. (2009). Proline-rich sequence recognition I: Marking GYF and WW domain assembly sites in early spliceosomal complexes. Mol. Cell. Proteom. 8 2461-2473.
Schlundt A, Sticht J, Piotukh K, Kosslick D, Jahnke N, Keller S, Schuemann M, Krause E, Freund C. (2009). Proline-rich sequence recognition II: proteomic analysis of TSG101-UEV interactions. Mol. Cell. Proteom. 8, 2474-2486.