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Interdomain contacts and the stability of serralysin protease from Serratia marcescens

Zhang, L and Morrison, AJ and Thibodeau, PH (2015) Interdomain contacts and the stability of serralysin protease from Serratia marcescens. PLoS ONE, 10 (9).

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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 Cterminal 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.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Zhang, Lliz46@pitt.eduLIZ46
Morrison, AJ
Thibodeau, PHthibodea@pitt.eduTHIBODEA
ContributionContributors NameEmailPitt UsernameORCID
Date: 17 September 2015
Date Type: Publication
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Journal or Publication Title: PLoS ONE
Volume: 10
Number: 9
DOI or Unique Handle: 10.1371/journal.pone.0138419
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Microbiology and Molecular Genetics
Refereed: Yes
Date Deposited: 23 Aug 2016 14:38
Last Modified: 30 Mar 2021 14:55


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