The solution structure of the soluble form of the lipid-modified azurin from Neisseria gonorrhoeae, the electron donor of cytochrome c peroxidase

• Published in: Biochimica et biophysica acta 2016-02, Vol.1857 (2), p.169-176
• Main Authors: Nóbrega, Cláudia S., Saraiva, Ivo H., Carreira, Cíntia, Devreese, Bart, Matzapetakis, Manolis, Pauleta, Sofia R.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Azurin; Azurin - chemistry; Azurin - genetics; Azurin - metabolism; Cloning, Molecular; Copper - chemistry; Copper - metabolism; Copper protein; Cytochrome c peroxidase; Cytochrome-c Peroxidase - chemistry; Cytochrome-c Peroxidase - genetics; Cytochrome-c Peroxidase - metabolism; Electron Transport; Electrons; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Hydrogen Peroxide - chemistry; Laz; Models, Molecular; Molecular Sequence Data; Neisseria; Neisseria gonorrhoeae - chemistry; Neisseria gonorrhoeae - enzymology; Oxidation-Reduction; Protein Folding; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Solution NMR structure; Substrate Specificity
• ISSN: 0005-2728; 0006-3002; 1879-2650
• DOI: 10.1016/j.bbabio.2015.11.006

Neisseria gonorrhoeae colonizes the genitourinary track, and in these environments, especially in the female host, the bacteria are subjected to low levels of oxygen, and reactive oxygen and nitrosyl species. Here, the biochemical characterization of N. gonorrhoeae Laz is presented, as well as, the solution structure of its soluble domain determined by NMR. N. gonorrhoeae Laz is a type 1 copper protein of the azurin-family based on its spectroscopic properties and structure, with a redox potential of 277±5mV, at pH7.0, that behaves as a monomer in solution. The globular Laz soluble domain adopts the Greek-key motif, with the copper center located at one end of the β-barrel coordinated by Gly48, His49, Cys113, His118 and Met122, in a distorted trigonal geometry. The edge of the His118 imidazole ring is water exposed, in a surface that is proposed to be involved in the interaction with its redox partners. The heterologously expressed Laz was shown to be a competent electron donor to N. gonorrhoeae cytochrome c peroxidase. This is an evidence for its involvement in the mechanism of protection against hydrogen peroxide generated by neighboring lactobacilli in the host environment. [Display omitted] •Neisseria gonorrhoeae Laz solution structure has the typical β-barrel fold.•Laz is a specific electron donor to Neisseria cytochrome c peroxidase.•Laz is a type 1 copper protein by visible and EPR spectroscopies.•Neisseria gonorrhoeae Laz can be heterologously produced in the holo-form.