Architecture and dynamics of macromolecular machines involved in homeostasis

• Applicant: Professor Dr. Florian Stengel
• Subject Area: Biochemistry ; Structural Biology
• Term: since 2015
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 282309713

Virtually all biological processes are controlled and catalysed by protein complexes and there has been great progress over the last few years in our understanding of the architecture of many of these large molecular machines. However, our knowledge of how these complexes are regulated on the level of the intact protein complex, is still highly rudimentary and to monitor the assembly state, spatial-temporal distribution and interaction of even the most abundant and ubiquitous protein complexes remains a significant challenge.Disentangling this interplay is key for understanding the regulation of protein maintenance, or proteome homeostasis, whose dysregulation is involved in a multitude of diseases. Here, a single high-resolution structural approach is unlikely to succeed. Rather, the combination and integration of multiple hybrid methods will be needed to interpret the interplay of such complex macromolecular assemblies. In this application, I propose to develop and apply novel mass spectrometric and proteomic approaches together with integrated modelling and genomic approaches to quantitatively study the content, assembly, interactions and dynamics of protein complexes involved in homeostasis.In particular, I will i) elucidate the architecture of core components of the translational machinery, ii) expand the structural mass spectrometry toolbox by enabling quantitative and dynamical probing of intact protein complexes in their cellular environment, and iii) decode the structural and spatial interaction landscape of macromolecular machines involved in homeostasis.The hybrid structural mass spectrometry (MS) based approach put forward in this proposal will not only allow me to address these fundamental questions of biology on the level of the intact complex, but will close a gap between high-resolution structural approaches on the level of the single protein complex and systems efforts based on quantitation of single proteins or peptides. As such it holds great promise to significantly increase our general understanding of hyper-macromolecular protein assemblies in the context of the cell and to revolutionize our understanding of key functional modules within the cell.

• DFG Programme: Independent Junior Research Groups
• Major Instrumentation: HPLC
• Instrumentation Group: 1350 Flüssigkeits-Chromatographen (außer Aminosäureanalysatoren 317), Ionenaustauscher