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GBMSDG Meeting Archives

April 27, 2011

Guest Speaker
Dr. Bernd Wollscheid
Institute of Molecular Systems Biology (IMSB)
NCCR Neuro Center for Proteomics
ETH Zurich Switzerland

Caught in the act: Ligand-based receptor capturing (LRC) on living cells

Location
Novartis Institutes for Biomedical Research (NIBR)
250 Massachusetts Avenue
Cambridge, MA

Sponsored by
Agilent Technologies

Special thanks to Novartis Institute for Biomdical Research for sponsoring and hostong this event. It is the active participation of our sponsors that allows us to keep the fees for membership, dinners, drinks, speakers, etc. so low.

Presentation Abstract

Ligand-induced changes in cell surface receptors result in physiological responses, which constitute the biological activity of various ligands such as proteins, peptides, pharmaceutical drugs, toxins or whole pathogens. However, traditional approaches for the ligand-based identification of corresponding receptors are usually limited to non-transient, high affinity interactions and highly artificial experimental set-ups. Therefore, many signaling molecules remain orphan ligands without a known primary molecular target – invaluable information in understanding the respective mechanisms of signal transduction, drug action or disease.

Previously, we have developed the cell surface capturing (CSC) technology for the unbiased identification and quantification of cell surface N-glycoproteomes by mass spectrometry (MS), sets of SRM assays for selected N-glycopeptides of clinical interest and the Cell Surface Protein Atlas (CSPA). This demonstrated the powerful applicability of chemical reagents in the tagging of cell surface glycoproteins at carbohydrate groups and the subsequent purification of the corresponding peptides for MS analysis.

Based on these results we now synthesized trifunctional cross-linkers for the ligand-based tagging of glycoprotein receptors on living cells and the purification of receptor-derived peptides for MS analysis. Through quantitative comparison to a sample generated with an unspecific control probe, this ligand-based receptor capturing (LRC) approach allows for the highly specific and sensitive detection of ligand interactions with their corresponding receptors under near-physiological conditions. Experiments with ligands ranging from peptide hormones to clinical antibodies demonstrate the potential of this approach to specifically identify one or more target receptors for a given ligand with great statistical power. Advanced discovery-driven applications reveal potential receptors and receptor panels for ligands ranging from protein domains to intact viruses.

Together, I will present a summary of our recent research to understand the surfaceome as a cellular information gateway and a chemoproteomic technology for the unbiased detection of ligand-receptor interactions on living cells.

About Dr. Wollscheid

Research focus: Surfaceome Biology; Development of a qualitative and quantitative understanding of the cell surface subproteome as a complex information gateway to the microenvironment.