Investigation of protein degradation mechanisms in Synechococcus elongates UTEX 2973
EMSL Project ID
49491
Abstract
The recently identified and characterized unicellular cyanobacteria, Synechococcus elongates UTEX 2973, a fast growing and genetically malleable organism, has the potential to become the definitive chassis for industrial production of biotechnology relevant products. We have begun to investigate the degradation of the light harvesting components in UTEX 2973 where the regulation and mechanism of cellular degradation has been largely understudied. A greater understanding of protein degradation will provide a fuller understanding of protein stability, and hence maximizing potential bio-product yields. UTEX 2973 provides an excellent model for such investigations, as experiments can be designed within the timescale of hours instead of days. Preliminary studies utilizing EMSLs top-down mass spectrometry has provided proof of principle identifications, where key phycobilisome protein components have been identified with chromophores and other relevant adducts, as well as the first proteomic identification of NblA, a small protein required for PBS degradation. Current needs include a more refined quantitative analysis of the phycobilisome complex in cellular degradation mode for precise chromophore localization, degradation product quantification, and isoform characterization of NblA. Limited UTEX 2973 test samples run on the high resolution Orbitrap Lumos Fusion hybrid Mass Spectrometer have verified the potential benefits of this platform for this characterization. Requested is the instrument time required to run 32 top-down analyses on the optimized Lumos Fusion along with the subsequent sample preparation and data analysis support. This data will augment already collected orthogonal physiological data of UTEX 2973 for a comprehensive characterization of the protein degradation process in cyanobacteria.
Project Details
Start Date
2016-06-09
End Date
2016-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members