Decoding Brain Complexity: Spatial Proteomics and Protein Turnover in Neurons

Prof. Eugenio Fornasiero - Georg-August-Universität Göttingen
Decoding Brain Complexity: Spatial Proteomics and Protein Turnover in Neurons

Abstract
Mapping the spatial structure and dynamic regulation of proteins at high resolution is required to comprehend brain function. We outline two complementary approaches to overcome these obstacles. First, SUM-PAINT (Secondary label based Unlimited Multiplexed DNA-PAINT) enables virtuallyunlimited multiplexed imaging at the nanometer scale, enabling single-molecule resolved spatial proteomics in neurons. The technique revealed synaptic heterogeneity on an unprecedented scale, disclosing various types of synapses and enabling integrated acquisition and analysis pipelines for high-content spatial proteomics. Second, Turnover-PPT, a proteomics approach, plots the abundance and lifetime of 11,000 proteins and 40,000 phosphosites across various tissues and brain subregions. Integrating stable isotope labeling with mass spectrometry, Turnover-PPT reveals protein turnover dynamics in brain subregions, which unveil the regulatory principles for protein stability and phosphorylation. Some of the key results are the engagement of phosphorylation-dependent stability of neurodegeneration-related proteins such as Tau and α synuclein. Collectively, these developments give new insights into protein spatial distribution and dynamic turnover and clarify general mechanisms of synaptic organization, protein homeostasis, and neurodegenerative disease processes. This seminar will discuss how cutting edge imaging and proteomics strategies can be used to crack the molecular complexity of the brain in health and disease.