Structure and proposed mechanism of α-glycerophosphate oxidase from Mycoplasma pneumoniae
Abstract
In this study, the formation of hydrogen peroxide (H₂O₂) by the FAD-dependent α-glycerophosphate oxidase (GlpO), is important for the pathogenesis of Streptococcus pneumoniae and Mycoplasma pneumoniae. The structurally known GlpO from Streptococcus sp. (SspGlpO) is similar to the pneumococcal protein (SpGlpO) and provides a guide for drug design against that target. However, M. pneumoniae GlpO (MpGlpO), having <20% sequence identity with structurally known GlpOs, appears to represent a second type of GlpO we designate as Type II GlpOs. Here, the recombinant His-tagged MpGlpO structure is described at ~2.5 Å resolution, solved by molecular replacement using as a search model the Bordetella pertussis protein 3253 (Bp3253) a protein of unknown function solved by structural genomics efforts. Recombinant MpGlpO is an active oxidase with a turnover number of ~580 min⁻¹ while Bp3253 showed no GlpO activity. No substantial differences exist between the oxidized and dithionite-reduced MpGlpO structures. Although, no liganded structures were determined, a comparison with the tartrate-bound Bp3253 structure and consideration of residue conservation patterns guided the construction of a model for α-glycerophosphate (Glp) recognition and turnover by MpGlpO. The predicted binding mode also appears relevant for the type I GlpOs (such as SspGlpO) despite differences in substrate recognition residues, andmore »
- Authors:
-
- Oregon State Univ., Corvallis, OR (United States)
- Center for Structural Biology, Wake Forest School of Medicine, Winston-Salem NC USA
- Burapha Univ., Chonburi (Thailand)
- Mahidol Univ., Bangkok (Thailand)
- Wake Forest School of Medicine, Winston-Salem, NC (United States)
- Publication Date:
- Research Org.:
- Oregon State Univ., Corvallis, OR (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1240120
- Grant/Contract Number:
- AC02-98CH10886
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Federation of European Biochemical Societies (FEBS) Journal
- Additional Journal Information:
- Journal Name: Federation of European Biochemical Societies (FEBS) Journal; Journal Volume: 282; Journal Issue: 16; Journal ID: ISSN 1742-464X
- Publisher:
- Federation of European Biochemical Societies
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Elkhal, Callia K., Kean, Kelsey M., Parsonage, Derek, Maenpuen, Somchart, Chaiyen, Pimchai, Claiborne, Al, and Karplus, P. Andrew. Structure and proposed mechanism of α-glycerophosphate oxidase from Mycoplasma pneumoniae. United States: N. p., 2015.
Web. doi:10.1111/febs.13233.
Elkhal, Callia K., Kean, Kelsey M., Parsonage, Derek, Maenpuen, Somchart, Chaiyen, Pimchai, Claiborne, Al, & Karplus, P. Andrew. Structure and proposed mechanism of α-glycerophosphate oxidase from Mycoplasma pneumoniae. United States. https://doi.org/10.1111/febs.13233
Elkhal, Callia K., Kean, Kelsey M., Parsonage, Derek, Maenpuen, Somchart, Chaiyen, Pimchai, Claiborne, Al, and Karplus, P. Andrew. Sat .
"Structure and proposed mechanism of α-glycerophosphate oxidase from Mycoplasma pneumoniae". United States. https://doi.org/10.1111/febs.13233. https://www.osti.gov/servlets/purl/1240120.
@article{osti_1240120,
title = {Structure and proposed mechanism of α-glycerophosphate oxidase from Mycoplasma pneumoniae},
author = {Elkhal, Callia K. and Kean, Kelsey M. and Parsonage, Derek and Maenpuen, Somchart and Chaiyen, Pimchai and Claiborne, Al and Karplus, P. Andrew},
abstractNote = {In this study, the formation of hydrogen peroxide (H₂O₂) by the FAD-dependent α-glycerophosphate oxidase (GlpO), is important for the pathogenesis of Streptococcus pneumoniae and Mycoplasma pneumoniae. The structurally known GlpO from Streptococcus sp. (SspGlpO) is similar to the pneumococcal protein (SpGlpO) and provides a guide for drug design against that target. However, M. pneumoniae GlpO (MpGlpO), having <20% sequence identity with structurally known GlpOs, appears to represent a second type of GlpO we designate as Type II GlpOs. Here, the recombinant His-tagged MpGlpO structure is described at ~2.5 Å resolution, solved by molecular replacement using as a search model the Bordetella pertussis protein 3253 (Bp3253) a protein of unknown function solved by structural genomics efforts. Recombinant MpGlpO is an active oxidase with a turnover number of ~580 min⁻¹ while Bp3253 showed no GlpO activity. No substantial differences exist between the oxidized and dithionite-reduced MpGlpO structures. Although, no liganded structures were determined, a comparison with the tartrate-bound Bp3253 structure and consideration of residue conservation patterns guided the construction of a model for α-glycerophosphate (Glp) recognition and turnover by MpGlpO. The predicted binding mode also appears relevant for the type I GlpOs (such as SspGlpO) despite differences in substrate recognition residues, and it implicates a histidine conserved in type I and II Glp oxidases and dehydrogenases as the catalytic acid/base. This work provides a solid foundation for guiding further studies of the mitochondrial Glp dehydrogenases as well as for continued studies of M. pneumoniae and S. pneumoniae glycerol metabolism and the development of novel therapeutics targeting MpGlpO and SpGlpO.},
doi = {10.1111/febs.13233},
journal = {Federation of European Biochemical Societies (FEBS) Journal},
number = 16,
volume = 282,
place = {United States},
year = {Sat Mar 14 00:00:00 EDT 2015},
month = {Sat Mar 14 00:00:00 EDT 2015}
}
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Works referencing / citing this record:
Potential Molecular Targets for Narrow-Spectrum Agents to Combat Mycoplasma pneumoniae Infection and Disease
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