Impact of selected amino acids of HP0377 (Helicobacter pylori thiol oxidoreductase) on its functioning as a CcmG (cytochrome c maturation) protein and Dsb (disulfide bond) isomerase

Magdalena Joanna Grzeszczuk , Aleksandra Bąk , Anna Marta Banaś , Paweł Urbanowicz , Stanislaw Dunin-Horkawicz , Artur Giełdoń , Cezary Czaplewski , Józef Adam Liwo , Elżbieta K. Jagusztyn-Krynicka

Abstract

Helicobacter pylori HP0377 is a thiol oxidoreductase, a member of the CcmG family involved in cytochrome biogenesis, as previously shown by in vitro experiments. In this report, we document that HP0377 also acts in vivo in the cytochrome assembly process in Bacillus subtilis, where it complements the lack of ResA. However, unlike other characterized proteins in this family, HP0377 is a dithiol reductase and isomerase. We elucidated how the amino acid composition of its active site modulates its functionality. We demonstrated that cis-proline (P156) is involved in its interaction with the redox partner (CcdA), as a P156T HP0377 variant is inactive in vivo and is present in the oxidized form in B. subtilis. Furthermore, we showed that engineering the HP0377 active motif by changing CSYC motif into CSYS or SSYC, clearly diminishes two activities (reduction and isomerization) of the protein. Whereas HP0377CSYA is inactive in reduction as well as in isomerization, HP0377CSYS retains reductive activity. Also, replacement of F95 by Q decreases its ability to regenerate scRNase and does not influence the reductive activity of HP0377CSYS towards apocytochrome c. HP0377 is also distinguished from other CcmGs as it forms a 2:1 complex with apocytochrome c. Phylogenetic analyses showed that, although HP0377 is capable of complementing ResA in Bacillus subtilis, its thioredoxin domain has a different origin, presumably common to DsbC.
Author Magdalena Joanna Grzeszczuk - [Uniwersytet Warszawski]
Magdalena Joanna Grzeszczuk,,
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, Aleksandra Bąk - [Uniwersytet Warszawski]
Aleksandra Bąk,,
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, Anna Marta Banaś - [Uniwersytet Warszawski]
Anna Marta Banaś,,
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, Paweł Urbanowicz - [Uniwersytet Warszawski]
Paweł Urbanowicz,,
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, Stanislaw Dunin-Horkawicz - [Laboratory of Structural Bioinformatics]
Stanislaw Dunin-Horkawicz,,
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, Artur Giełdoń (FCh / DTCh / LSP)
Artur Giełdoń,,
- Laboratory of Simulation of Polymers
, Cezary Czaplewski (FCh / DTCh / LSP)
Cezary Czaplewski,,
- Laboratory of Simulation of Polymers
, Józef Adam Liwo (FCh / DTCh / LMM)
Józef Adam Liwo,,
- Laboratory of Molecular Modeling
, Elżbieta K. Jagusztyn-Krynicka - [Uniwersytet Warszawski]
Elżbieta K. Jagusztyn-Krynicka,,
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Journal seriesPLoS ONE, ISSN 1932-6203, (A 40 pkt)
Issue year2018
Vol13
No4
Pages1-27
Publication size in sheets1.3
Keywords in Englishextracytoplasmic thioredoxin resa; bacillus-subtilis resa; bradyrhizobium-japonicum; substrate-specificity; cysteine residues; crystal-structure; secretion system; structural basis; biogenesis; redox
ASJC Classification2700 General Medicine; 1100 General Agricultural and Biological Sciences; 1300 General Biochemistry, Genetics and Molecular Biology
DOIDOI:10.1371/journal.pone.0195358
URL https://doi.org/10.1371/journal.pone.0195358
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)40
Score sourcejournalList
ScoreMinisterial score = 40.0, 20-03-2020, ArticleFromJournal
Publication indicators WoS Citations = 0; Scopus Citations = 2; Scopus SNIP (Source Normalised Impact per Paper): 2017 = 1.111; WoS Impact Factor: 2018 = 2.776 (2) - 2018=3.337 (5)
Citation count*2 (2020-03-21)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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