Confident identification of citrullinated peptides from P gingivalis

ABSTRACT: Porphyromonas gingivalis is a key pathogen in the development of chronic periodontitis and has recently been linked to the development of rheumatoid arthritis and Alzheimer’s disease. In this project the outer membrane vesicles (OMV) from Pg have been analyzed using a two-dimensional HFBA-based separation system combined with LC-MS. Three strains from P. gingivalis was analyzed in order to elucidate their citrullinome: wild-type (WT), a mutant with the active site cysteine mutated to alanine (C351A), as well as a knock-out mutant of peptidyl arginine deiminase (ΔPPAD). For optimal and positive identification and validation of citrullinated peptides and proteins, high resolution mass spectrometers and strict MS criteria were utilized. This may have compromised the total number of identified citrullination, but increased the confidence of the validation. A new 2D separation system proved to increase the strength of validation, and along with the use of an in-house build program, Citrullia, we establish a fast and easy semi-automatic (manual) validation of citrullinated peptides. For WT we identified 78 citrullinated proteins having a total of 161 citrullination sites. For C351A a single citrullination site was found and no citrullinations was found for ΔPPAD.

INSTRUMENT(S):

ORGANISM(S): Porphyromonas Gingivalis (strain Atcc Baa-308 / W83)

SUBMITTER: Peter Højrup  

LAB HEAD: Peter Højrup

PROVIDER: PXD015701 | Pride | 2019-11-22

REPOSITORIES: pride