Hydrogen peroxide and hypochlorous acid influx through the major S. Typhimurium porin OmpD is affected by substitution of key residues of the channel

• Published in: Archives of biochemistry and biophysics 2015-02, Vol.568, p.38-45
• Main Authors: Aguayo, Daniel, Pacheco, Nicolás, Morales, Eduardo H., Collao, Bernardo, Luraschi, Roberto, Cabezas, Carolina, Calderón, Paulina, González-Nilo, Fernando, Gil, Fernando, Calderón, Iván L., Saavedra, Claudia P.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino Acid Substitution; Homology modeling; Hydrogen peroxide; Hydrogen Peroxide - metabolism; Hypochlorous acid; Hypochlorous Acid - metabolism; Molecular Dynamics Simulation; Molecular Sequence Data; Molecular simulations; OmpD porin; Porins - chemistry; Porins - genetics; Porins - metabolism; Salmonella typhimurium - chemistry; Salmonella typhimurium - genetics; Salmonella typhimurium - metabolism; Sequence Alignment
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2015.01.005

[Display omitted] •The S. Typhimurium OmpD porin mediates H2O2 and NaOCl transport.•Substitution of key residues of the OmpD channel modifies H2O2 and NaOCl transport through the porin.•Substitution of key residues of the OmpD channel increases S. Typhimurium resistance to H2O2 and NaOCl. OmpD is the major Salmonella enterica serovar Typhimurium (S. Typhimurium) porin and mediates hydrogen peroxide (H2O2) influx. The results described herein extend this finding to hypochlorous acid (HOCl), another reactive oxygen species that is also part of the oxidative burst generated by the phagosome. S. Typhimurium cells lacking OmpD show decreased HOCl influx, and OmpD-reconstituted proteoliposomes show an increase in the uptake of the toxic compound. To understand this physiologically relevant process, we investigated the role of key OmpD residues in H2O2 and NaOCl transport. Using a theoretical approach, residue K16 was defined as a major contributor to the channel electrostatic properties, and E111 was shown to directly participate in the size-exclusion limit of the channel. Together, we provide theoretical, genetic, and biochemical evidence that OmpD mediates H2O2 and NaOCl uptake, and that key residues of the channel are implicated in this process.