PMID- 26378460
OWN - NLM
STAT- MEDLINE
DCOM- 20160520
LR  - 20201215
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 9
DP  - 2015
TI  - Interdomain Contacts and the Stability of Serralysin Protease from Serratia 
      marcescens.
PG  - e0138419
LID - 10.1371/journal.pone.0138419 [doi]
LID - e0138419
AB  - The serralysin family of bacterial metalloproteases is associated with virulence 
      in multiple modes of infection. These extracellular proteases are members of the 
      Repeats-in-ToXin (RTX) family of toxins and virulence factors, which mediated 
      virulence in E. coli, B. pertussis, and P. aeruginosa, as well as other animal 
      and plant pathogens. The serralysin proteases are structurally dynamic and their 
      folding is regulated by calcium binding to a C-terminal domain that defines the 
      RTX family of proteins. Previous studies have suggested that interactions between 
      N-terminal sequences and this C-terminal domain are important for the high 
      thermal and chemical stabilities of the RTX proteases. Extending from this, 
      stabilization of these interactions in the native structure may lead to 
      hyperstabilization of the folded protein. To test this hypothesis, cysteine pairs 
      were introduced into the N-terminal helix and the RTX domain and protease folding 
      and activity were assessed. Under stringent pH and temperature conditions, the 
      disulfide-bonded mutant showed increased protease activity and stability. This 
      activity was dependent on the redox environment of the refolding reaction and 
      could be blocked by selective modification of the cysteine residues before 
      protease refolding. These data demonstrate that the thermal and chemical 
      stability of these proteases is, in part, mediated by binding between the RTX 
      domain and the N-terminal helix and demonstrate that stabilization of this 
      interaction can further stabilize the active protease, leading to additional pH 
      and thermal tolerance.
FAU - Zhang, Liang
AU  - Zhang L
AD  - Department of Microbiology and Molecular Genetics, University of Pittsburgh, 
      School of Medicine, Pittsburgh, PA 15219, United States of America.
FAU - Morrison, Anneliese J
AU  - Morrison AJ
AD  - Department of Microbiology and Molecular Genetics, University of Pittsburgh, 
      School of Medicine, Pittsburgh, PA 15219, United States of America.
FAU - Thibodeau, Patrick H
AU  - Thibodeau PH
AD  - Department of Microbiology and Molecular Genetics, University of Pittsburgh, 
      School of Medicine, Pittsburgh, PA 15219, United States of America.
LA  - eng
GR  - R01 DK083284/DK/NIDDK NIH HHS/United States
GR  - R56 DK083284/DK/NIDDK NIH HHS/United States
GR  - T32 GM007759/GM/NIGMS NIH HHS/United States
GR  - DK083284/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20150917
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Virulence Factors)
RN  - EC 3.4.- (Peptide Hydrolases)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - EC 3.4.24.40 (serralysin)
SB  - IM
MH  - Metalloendopeptidases/*metabolism
MH  - Peptide Hydrolases/*metabolism
MH  - Protein Folding
MH  - Protein Structure, Secondary
MH  - Protein Structure, Tertiary
MH  - Serratia marcescens/*metabolism
MH  - Virulence Factors/metabolism
PMC - PMC4574703
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2015/09/18 06:00
MHDA- 2016/05/21 06:00
PMCR- 2015/09/17
CRDT- 2015/09/18 06:00
PHST- 2015/08/06 00:00 [received]
PHST- 2015/08/30 00:00 [accepted]
PHST- 2015/09/18 06:00 [entrez]
PHST- 2015/09/18 06:00 [pubmed]
PHST- 2016/05/21 06:00 [medline]
PHST- 2015/09/17 00:00 [pmc-release]
AID - PONE-D-15-34620 [pii]
AID - 10.1371/journal.pone.0138419 [doi]
PST - epublish
SO  - PLoS One. 2015 Sep 17;10(9):e0138419. doi: 10.1371/journal.pone.0138419. 
      eCollection 2015.