Exploring alternative proteases and fragmentation methods for proteomics — ASN Events
  1. Schedule
  2. Keynote Lecture
  3. Exploring alternative proteases and fragmentation methods for proteomics

Exploring alternative proteases and fragmentation methods for proteomics (#1)

Albert Heck 1
  1. Utrecht University, Utrecht, NL, Netherlands

Proteome analysis heavily relies on a broad mixture of analytical techniques, starting from sample preparation, to separation and enrichment and last but certainly not in the least mass spectrometry. Through developments in these enabling technologies MS-based proteomics has matured and start to deliver biological relevant information. Still our measurements are still far from comprehensive, and the optimal matured workflow does not (yet) exist for proteomics. In my laboratory we try to explore alternative complementary methods to explore better the richness of the proteome

In more detail I will describe methods enabling 1) enhanced peptide coverage based on a combination of HCD and ETD that leads to improved analysis of phosphopeptides, MHC peptides 1 and protein disulfide bridges 2, 2) enhanced protein and proteome coverage, by using multiple proteases and 3) improved analysis of intact proteins and protein complexes enabling to visualize all proteoforms of a given protein at once and following kinase phosphorylation in real-time using top-down approaches 3
  1. Mommen et al. PNAS 111 (2013) 4507-4512
  2. Liu et al. Mol Cell Prot 13 (2014) 2776-2786
  3. Van den Waterbeemd et al. Angew Chemie 53 (2014) 9660-9664