The GlycoPaSER Prototype as a Real-Time N-Glycopeptide Identification Tool Based on the PaSER Parallel Computing...

Area of Science:

• Proteomics
• Glycoproteomics
• Computational Biology

Background:

• Real-time database searching streamlines proteomics workflows and enables results-dependent acquisition (RDA).
• Glycopeptide analysis presents challenges due to diverse physicochemical properties requiring adaptable acquisition parameters.

Purpose of the Study:

• To establish the GlycoPaSER prototype for real-time glycopeptide identification.
• To leverage real-time analysis for results-dependent acquisition in glycoproteomics.

Main Methods:

• Extended the Parallel Search Engine in Real-time (PaSER) for glycopeptide analysis.
• Decomposed glycopeptide fragmentation spectra into peptide and glycan moieties.
• Employed specialized algorithms for real-time identification of each moiety.

Main Results:

• GlycoPaSER achieves real-time analysis, matching the pace of data acquisition.
• The prototype demonstrates comparable performance to existing glycoproteomics software.
• Results align with established literature reference data.

Conclusions:

• GlycoPaSER is the first proof-of-concept for real-time glycopeptide identification.
• This technology facilitates the future development of RDA for enhanced data acquisition.
• Real-time analysis significantly benefits complex glycoproteomics studies.