Development of a functional proteomics workflow for characterization of glycoproteins
EMSL Project ID
51671
Abstract
Glycosylation is a diverse family of post-translational modification (PTM) that affects protein structure/function but is not directly encoded in the genome, and represents significant challenge for existing omics approaches. Many plant membrane receptors for cellular signaling and secreted microbial enzymes relevant for biofuel production are heavily glycosylated. Despite the importance of glycosylation, current proteomics pipeline at EMSL is largely incapable of decoding the functional roles of glycoproteins. We propose to develop a robust high throughput workflow for glycoprotein characterization by implementing state-of-the-art separation and mass spectrometry techniques. This will provide "improved methods for charactering a diverse suite of post-translationally modified proteins" directly addressing EMSL Roadmap "DigiPhen" decadal goal. The workflow will produce mass spectrometry data at three levels - intact protein level for determining combinatorial PTMs, glycopeptide level for PTM site localization, and glycan level for confirming glycan identities. We will develop bioinformatics pipelines to integrate the data to precisely define the functional forms of glycoproteins. The established methods will be beta-tested on soybean and fungal laccase samples (provided by team members and collaborators) to reveal unique glycosylation patterns that can be associated with biological activity and phenotypes relevant to bioenergy applications.
Project Details
Start Date
2020-10-26
End Date
2021-11-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members