So grateful and honored to receive the MCP Lectureship Award at the #ASBMBProteomics meeting at the Broad Institute! Huge thanks to the wonderful organizers and to everyone—past and present—who’s been part of our lab.

3/3 We extended its application to crude human cell line, bacteria, and tissue lysates and sensitively identify membrane targets! It works extremely well in tissues which allowed us to identify off-targets for sunitinib in mouse heart tissue and could explain its well-known cardiotoxicity.

2/3 We provide a large collection of ATP-binding proteins in E.coli and give rich information on how they respond to ATP. Our data nicely showed how chaperon protein DnaK binds ATP at ATP-binding region at low ATP concentration and dissociates the substrate protein at high ATP concentration.

1/3 We extended the original PELSA protocol to work with 96-well plates and show a 100-fold higher throughput, excellent reproducibility, accurate determination of drug-binding affinities in dose-response experiments as shown by a strong correlation of 0.88 with kinobeads-based measurements.

Finally, we once again uncovered site-specific modulation of glycosylation upon perturbation, meaning that only some glycoforms on some glycosites are modulated, even within the same protein, suggesting complex regulatory mechanisms.

Next, we showed remodeling of the mouse brain glycoproteome upon gut microbiome colonization 🐭 The link b/w gut microbiome & brain physiology is long known, but molecular mechanisms remain elusive. We showed that proteins involved in neurotransmission & axon guidance were particularly affected.

We next used our quantitative approach to measure the dynamics of glycosylation changes in human cells treated with a fucosylation inhibitor 🧫 We discovered pervasive site-specific modulation of glycosylation upon perturbation.

Very little is known about glycoform functionality. We developed a functional glycoproteomics approach enabling the proteome-wide characterization of the solubility of different glycosylated proteoforms in the mouse brain 🐭

To understand structural features governing the level of site microheterogeneity, we leveraged our comprehensive dataset in combination with AlphaFold DB 🖥️

It is essential to differentiate surface-exposed glycoforms from intermediate species. To address this, we treated intact living human cells with two enzymes, enabling proteome-wide characterization of mature glycoforms 🧫

Using our quant approach, we discovered that many sites have similar glycosylation patterns across tissues 🐭However, we identified a subset of sites driving tissue-specificity, enriched in domains involved in cell adhesion, signaling & immunity. E.g., only N445 exhibits tissue specificity on INSR.

We are very happy to present our work on N-glycoproteomics!🍬 Our method enables the selective enrichment and precise quantification of intact N-glycopeptides to explore the dynamics of glycosylation microheterogeneity.
www.nature.com/articles/s41...