Extension of Species-Level Proteome Coverage in a Natural Microbial Community with the GELFrEE Fractionation System

摘要

FASP or TCA precipitation provides optimal clean up of proteins from GELFrEE buffer prior to digestion and 2D-LC-MS/MS. Fractionation of the C75 V2 biofilm resulted in increased proteomic coverage, using equivalent instrument time as unfractionated, when coupled with the LTQ Velos. GELFrEE fractionation and subcellular fractionation of ABFront revealed similar protein IDs with substantially less instrument time, and enabled the identification of 6,651 proteins representing the most proteins ever identified from a single AMD biofilm location. The next approach to increase proteomic coverage will combine subcellular fractionation with subsequent GELFrEE fractionation.