Z 1 Johannes Buchner Michael Sattler Lehrstuhl fr

Z 1 Johannes Buchner Michael Sattler Lehrstuhl für Biotechnologie Lehrstuhl für biomolekulare NMR-Spektroskopie Project Z 1: 'Biophysical Methods' Native and H/DX Mass Spectrometry Native MS of non-covalent protein complexes: Historical Example: Analysis of the s. Hsps Ta. Hsp 16. 9 Dynamics of the complex: Subunit exchange Complex stoichiometry Sobott et al. , Robinson, J. Biol Chem (2002) H/DX MS H/D-X MS: Experimental setup Hydrogen/Deuterium exchange ACQUITY UPLC M-class HDX with automation Digest Variation of reaction mixture/conditions D 2 O Quench reaction at various time points Intact protein time points UPLC separation Deuterium incorporation curves Data analysis ESI-TOF MS MS Automated system is necessary to enable reproducibility ! Engen et al. , Weis, Int Rev Phys Chem (2013) Example: Analysis of Src-kinase mutations Allosteric changes in the dynamics Peptide 162 -172: SH 2 -domain Peptide 270 -278: b 1 -strand & P-loop point mutation no significant differences: Mutation does not change dynamics of this region significant differences: Mutation changes dynamics of this region • Colors: increased H/DX increased dynamics • Data provide restraints for MD simulations Boczek et al. , Buchner, PNAS (2015) Support People dedicated Ralf Strehle Martin Haslbeck Gina Feind Ph. D students from project A 12 and A 03 User support MS Experiments & setup Service measurements Use of instruments Data analysis & evaluation Training courses Structures Baumeister: A 01 Groll: A 02, Conti: A 07 Mass spectrometry Sieber A 09, Hartl A 12 SAXS annual training course Native & H/DX MS SPR annual training course monthly discussion group Biophysical chemistry Buchner: A 03, A 06 Schwenkert: A 04 Organic chemistry Lang: B 10 Z 1 NMR Sattler: B 03, Hagn: B 13 Cell biology Feige: B 11 Further collaborations: Itzen: B 05, Kapurniotu B 06, Reif B 07