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Title: Top-Down Characterization of the Post-Translationally Modified Intact Periplasmic Proteome from the Bacterium Novosphingobium aromaticivorans

The periplasm of Gram-negative bacteria is a dynamic and physiologically important subcellular compartment where the constant exposure to potential environmental insults amplifies the need for proper protein folding and modifications. Top-down proteomics analysis of the periplasmic fraction at the intact protein level provides unrestricted characterization and annotation of the periplasmic proteome, including the post-translational modifications (PTMs) on these proteins. Here, we used single-dimension ultra-high pressure liquid chromatography coupled with the Fourier transform mass spectrometry (FTMS) to investigate the intact periplasmic proteome of Novosphingobium aromaticivorans . Our top-down analysis provided the confident identification of 55 proteins in the periplasm and characterized their PTMs including signal peptide removal, N-terminal methionine excision, acetylation, glutathionylation, pyroglutamate, and disulfide bond formation. This study provides the first experimental evidence for the expression and periplasmic localization of many hypothetical and uncharacterized proteins and the first unrestrictive, large-scale data on PTMs in the bacterial periplasm.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [1] ;  [1] ;  [5] ;  [3] ;  [3] ;  [3] ;  [3] ;  [1]
  1. Environmental Molecular Science Laboratory, Pacific Northwest National Laboratory, P.O. Box 999/MS K8-98, Richland, WA 99352, USA
  2. Center for Bioproducts and Bioenergy, Washington State University, Richland, WA, USA
  3. Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, USA
  4. Computational Sciences and Mathematics Division, Pacific Northwest National Laboratory, Richland, WA, USA
  5. Department of Neurobiology, 720 Westview Drive SW, Atlanta, GA, USA
Publication Date:
OSTI Identifier:
1197932
Grant/Contract Number:
AC05-76RLO-1830; 1097055
Type:
Published Article
Journal Name:
International Journal of Proteomics
Additional Journal Information:
Journal Volume: 2013; Related Information: CHORUS Timestamp: 2016-08-18 13:30:13; Journal ID: ISSN 2090-2166
Publisher:
Hindawi Publishing Corporation
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
Country unknown/Code not available
Language:
English