Mapping Cellular Microenvironments – Proximity Labeling/MS within a Campus Core Facility Structure (GoeCoOp-PL/MS)

• Subject Area: Metabolism, Biochemistry and Genetics of Microorganisms ; Cell Biology
• Term: since 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 502174128

Proximity-dependent labeling with biotin in living cells followed by mass spectrometry analysis (PL/MS) advances to a key technology to study protein-protein interactions and molecular microenvironments (called Biotin IDentification, BioID). Genetically engineered biotin ligases and split variants thereof (Split-BioID) with improved versatility and speed are currently developed to enable the analysis of context-specific and short-lived protein-protein interactions. Demanding quantitative mass spectrometry approaches that consider appropriate control cell lines and that apply proven workflows for MS data analysis are prerequisites for high quality results generated with this methodology. The Core Facility Protein Analytics (CF-PA) did pioneering work with PL/MS in Saccharomyces cerevisiae resulting in a model study for the proximity labeling of a ribosomal microenvironment. Currently the application of this methodology is broadened within the CF-PA to further fungal and microbial organisms and plants. An open PL/MS service module named GoeCoOp-PL/MS will be provided on the basis of close collaboration with Campus colleagues, e.g. members of the Goettingen Proteomics Forum (GPF), and external scientists to boost the local expertise for this fundamental technology. Furthermore, the possibilities of proximity labeling of metabolites with biotin and MS analysis will be studied and tested for routine applicability.

• DFG Programme: Core Facilities
• Applicant Institution: Georg-August-Universität Göttingen
• Leader: Dr. Oliver Valerius