Helicobacter pylori HP0377, a member of the Dsb family, is an untypical multifunctional CcmG that cooperates with dimeric thioldisulfide oxidase HP0231

BMC Microbiology

E. Katarzyna Jagusztyn-Krynicka²  Elzbieta Nowak¹  Piotr Wincek²  Paweł Urbanowicz²  Ewa Wywial³  Patrycja Kobierecka²  Magdalena Grzeszczuk²  Paula Roszczenko⁴ 

[1] Laboratory of Protein Structure, International Institute of Molecular and Cell Biology, Warsaw, Poland;Department of Bacterial Genetics, Institute of Microbiology, Faculty of Biology, University of Warsaw, Warsaw, Poland;Laboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology, Warsaw, Poland;Present address: Department of Cell Biology, Nencki Institute of Experimental Biology, PAS, Warsaw, Poland

关键词: Isomerase activity;    Cytochrome c biogenesis;    Helicobacter pylori;    Dsb;   

Others  :  1227628
DOI  :  10.1186/s12866-015-0471-z

 received in 2015-02-16, accepted in 2015-06-23,  发布年份 2015

【 摘 要 】

Background

In the genome of H. pylori 26695, 149 proteins containing the CXXC motif characteristic of thioldisulfide oxidoreductases have been identified to date. However, only two of these proteins have a thioredoxin-like fold (i.e., HP0377 and HP0231) and are periplasm-located. We have previously shown that HP0231 is a dimeric oxidoreductase that catalyzes disulfide bond formation in the periplasm. Although HP0377 was originally described as DsbC homologue, its resolved structure and location of the hp0377 gene in the genome indicate that it is a counterpart of CcmG/DsbE.

Results

The present work shows that HP0377 is present in H. pylori cells only in a reduced form and that absence of the main periplasmic oxidase HP0231 influences its redox state. Our biochemical analysis indicates that HP0377 is a specific reductase, as it does not reduce insulin. However, it possesses disulfide isomerase activity, as it catalyzes the refolding of scrambled RNase. Additionally, although its standard redox potential is -176 mV, it is the first described CcmG protein having an acidic pKa of the N-terminal cysteine of the CXXC motif, similar to E. coli DsbA or E. coli DsbC. The CcmG proteins that play a role in a cytochrome c-maturation, both in system I and system II, are kept in the reduced form by an integral membrane protein DsbD or its analogue, CcdA. In H. pylori HP0377 is re-reduced by CcdA (HP0265); however in E. coli it remains in the oxidized state as it does not interact with E. coli DsbD. Our in vivo work also suggests that both HP0377, which plays a role in apocytochrome reduction, and HP0378, which is involved in heme transport and its ligation into apocytochrome, provide essential functions in H. pylori.

Conclusions

The present data, in combination with the resolved three-dimensional structure of the HP0377, suggest that HP0377 is an unusual, multifunctional CcmG protein.

【 授权许可】

   
2015 Roszczenko et al.

【 图 表 】

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