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Characterization of S-thiolation on secreted proteins from E. coli by mass spectrometry.

Authors
  • Liu, Peiran
  • Tarnowski, Malloree A
  • O'Mara, Brian W
  • Wu, Wei
  • Zhang, Haiying
  • Tamura, James K
  • Ackerman, Michael S
  • Tao, Li
  • Grace, Michael J
  • Russell, Reb J
Type
Published Article
Journal
Rapid Communications in Mass Spectrometry
Publisher
Wiley (John Wiley & Sons)
Publication Date
Oct 30, 2009
Volume
23
Issue
20
Pages
3343–3349
Identifiers
DOI: 10.1002/rcm.4247
PMID: 19760645
Source
Medline
License
Unknown

Abstract

S-thiolation is a reversible post-translational modification in which thiol metabolites of low molecular masses are linked to protein sulfhydryl groups through disulfide bonds. This modification is commonly observed in recombinant proteins secreted from E. coli cells. Since it can alter protein functions and introduce molecular heterogeneity, S-thiolation is undesirable for recombinant protein production. To date, few published studies have characterized thiol modifiers or investigated the mechanism of S-thiolation in recombinant proteins. In this work, reversed-phase liquid chromatography and mass spectrometry were used to characterize four of the most abundant thiol modifiers on recombinant proteins secreted from E. coli BL21 (DE3) strain. These thiol modifiers have been identified as glutathione, 4-phosphopantetheine, gluconoylated glutathione, and dephosphorylated coenzyme A. S-thiolation by these thiol modifiers increases protein mass by 305, 356, 483, and 685 Da, respectively. These specific mass increases can be used as markers for identifying S-thiolation in recombinant proteins.

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