These authors contributed equally to this work
DegP primarily functions as a protease for the biogenesis of β-barrel outer membrane proteins in the Gram-negative bacterium Escherichia coli
Article first published online: 15 JAN 2014
© 2013 FEBS
Volume 281, Issue 4, pages 1226–1240, February 2014
How to Cite
Ge, X., Wang, R., Ma, J., Liu, Y., Ezemaduka, A. N., Chen, P. R., Fu, X. and Chang, Z. (2014), DegP primarily functions as a protease for the biogenesis of β-barrel outer membrane proteins in the Gram-negative bacterium Escherichia coli. FEBS Journal, 281: 1226–1240. doi: 10.1111/febs.12701
- Issue published online: 17 FEB 2014
- Article first published online: 15 JAN 2014
- Accepted manuscript online: 26 DEC 2013 07:41AM EST
- Manuscript Accepted: 17 DEC 2013
- Manuscript Revised: 16 DEC 2013
- Manuscript Received: 22 AUG 2013
- National Basic Research Program of China. Grant Number: 2012CB917300
- National Natural Science Foundation of China. Grant Numbers: 31170738, 31100559, 31270804
Fig. S1. Immunoblotting results, using the antibody against His-tag, of photo-crosslinked protein products of DegP(S210A)His6 variants after being separated by SDS/PAGE.
Fig. S2. Specificities of the anti-OMP antibodies used in this study.
Fig. S3. Coomassie Blue staining results of proteins co-purified with DegP(S210A).
Fig. S4. The OmpC, OmpF and the transmembrane domain of OmpA were captured by DegP(S210A) largely in their unfolded states.
Fig. S5. The degP− cells cultured at 44 °C for 6 h are still alive.
Fig. S6. Misfolded OMPs already accumulate in the degP− cells under sub-lethal conditions.
Fig. S7. Chaperone function of SurA and protease function of DegP in OMP biogenesis.
Fig. S8. Coomassie Blue staining results of purified SurA, DegP, DegP(S210A) and OmpF, as well as the isolated outer membrane fraction (lacking OmpC and OmpF; at position indicated by the arrow) used in the in vitro refolding/reassembly assay.
Fig. S9. The membrane integrity is impaired in the degP− mutant cells cultured at 44 °C.
Fig. S10. Overexpressed DegP(S210A) prevents the toxic accumulation of misfolded OMPs onto the outer membrane but does not refold them.
Doc. S1. Experimental procedures.
Table S1. Abundance of OMPs and other representative envelope proteins in E. coli cells.
Table S2. Relation between misfolded OMPs and growth defect of strains analyzed in this study.
Table S3. E. coli strains used in this study.
Table S4. Plasmids used in this study.
Table S5. Primers used in this study.
Table S6. Antibodies used in this study.
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