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Biophysical and Biochemical Characterization of TP0037, a D-Lactate Dehydrogenase, Supports an Acetogenic Energy Conservation Pathway in Treponema pallidum

Journal Article · · mBio (Online)
 [1];  [1];  [1];  [1]
  1. UT Southwestern Medical Center, Dallas, TX (United States)
+ Show Author Affiliations
A longstanding conundrum in Treponema pallidum biology concerns how the spirochete generates sufficient energy to fulfill its complex pathogenesis processes during human syphilitic infection. For decades, it has been assumed that the bacterium relies solely on glucose catabolism (via glycolysis) for generation of its ATP. However, the organism’s robust motility, believed to be essential for human tissue invasion and dissemination, would require abundant ATP likely not provided by the parsimony of glycolysis. As such, additional ATP generation, either via a chemiosmotic gradient, substrate-level phosphorylation, or both, likely exists in T. pallidum. Along these lines, we have hypothesized that T. pallidum exploits an acetogenic energy conservation pathway that relies on the redox chemistry of flavins. Central to this hypothesis is the apparent existence in T. pallidum of an acetogenic pathway for the conversion of d-lactate to acetate. Herein we have characterized the structural, biophysical, and biochemical properties of the first enzyme (d-lactate dehydrogenase [d-LDH]; TP0037) predicted in this pathway. Binding and enzymatic studies showed that recombinant TP0037 consumed d-lactate and NAD+ to produce pyruvate and NADH. The crystal structure of TP0037 revealed a fold similar to that of other d-acid dehydrogenases; residues in the cofactor-binding and active sites were homologous to those of other known d-LDHs. The crystal structure and solution biophysical experiments revealed the protein’s propensity to dimerize, akin to other d-LDHs. This study is the first to elucidate the enzymatic properties of T. pallidum’s d-LDH, thereby providing new compelling evidence for a flavin-dependent acetogenic energy conservation (ATP-generating) pathway in T. pallidum.
Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Contributing Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1729697
Journal Information:
mBio (Online), Journal Name: mBio (Online) Journal Issue: 5 Vol. 11; ISSN 2150-7511
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

References (81)

The monocarboxylate transporter family-Role and regulation journal December 2011
Molecular insights into the enzymatic diversity of flavin-trafficking protein (Ftp; formerly ApbE) in flavoprotein biogenesis in the bacterial periplasm journal December 2015
Insights into the potential function and membrane organization of the TP0435 (Tp17) lipoprotein from T reponema pallidum derived from structural and biophysical analyses: Disulfide-Linked Dimer of T. pallidum TP0435 journal October 2014
Functional clues from the crystal structure of an orphan periplasmic ligand-binding protein from Treponema pallidum : Tp0737 Protein of Treponema Pallidum journal March 2017
Crystal stuctures of MglB-2 (TP0684), a topologically variant d -glucose-binding protein from Treponema pallidum, reveal a ligand-induced conformational change: Conformational Changes in TpMglB-2 journal February 2018
Lactate oxidation byTreponema pallidum journal May 1986
The sodium pumping NADH:quinone oxidoreductase (Na+-NQR), a unique redox-driven ion pump journal July 2014
Anatomical and biochemical evidence for Treponema pallidum in a 19th to early twentieth century skeletal cadaver journal May 2020
Cloning and overexpression of the Lactobacillus bulgaricus NAD+-dependent D-lactate dehydrogenase gene in Escherichia coli: Purification and characterization of the recombinant enzyme journal June 1992
Properties of ?-lactate dehydrogenase from Lactobacillus bulgaricus: a possible different evolutionary origin for the ?- and ?-lactate dehydrogenases journal March 1991
Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation Modeling journal March 2000
Size-Distribution Analysis of Proteins by Analytical Ultracentrifugation: Strategies and Application to Model Systems journal February 2002
Domain Closure, Substrate Specificity and Catalysis of d-Lactate Dehydrogenase from Lactobacillus bulgaricus journal April 2002
[20] Processing of X-ray diffraction data collected in oscillation mode book January 1997
Calculations and Publication-Quality Illustrations for Analytical Ultracentrifugation Data book January 2015
Sodium-translocating NADH:quinone oxidoreductase as a redox-driven ion pump journal June 2010
Inference of Macromolecular Assemblies from Crystalline State journal September 2007
Structural and Biochemical Basis for Polyamine Binding to the Tp0655 Lipoprotein of Treponema pallidum: Putative Role for Tp0655 (TpPotD) as a Polyamine Receptor journal October 2007
Cellular Architecture of Treponema pallidum: Novel Flagellum, Periplasmic Cone, and Cell Envelope as Revealed by Cryo Electron Tomography journal November 2010
Structural, Bioinformatic, and In Vivo Analyses of Two Treponema pallidum Lipoproteins Reveal a Unique TRAP Transporter journal March 2012
Structural and Thermodynamic Characterization of the Interaction between Two Periplasmic Treponema pallidum Lipoproteins that are Components of a TPR-Protein-Associated TRAP Transporter (TPAT) journal June 2012
Predicting the ligand-binding properties of Borrelia burgdorferi s.s. Bmp proteins in light of the conserved features of related Borrelia proteins journal February 2019
Insights into substrate binding by D-2-ketoacid dehydrogenases from the structure of Lactobacillus pentosus D-lactate dehydrogenase journal April 1996
D-lactic acidosis: Turning sugar into acids in the gastrointestinal tract journal January 1996
Structure of the V. cholerae Na+-pumping NADH:quinone oxidoreductase journal December 2014
The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability journal September 2007
Automated macromolecular model building for X-ray crystallography using ARP/wARP version 7 journal June 2008
Syphilis journal October 2017
Treponema pallidum, the syphilis spirochete: making a living as a stealth pathogen journal October 2016
Flavin transferase: the maturation factor of flavin-containing oxidoreductases journal August 2018
Energy conservation involving 2 respiratory circuits journal December 2019
The TP0796 Lipoprotein of Treponema pallidum Is a Bimetal-dependent FAD Pyrophosphatase with a Potential Role in Flavin Homeostasis journal February 2013
Reduction of Flavodoxin by Electron Bifurcation and Sodium Ion-dependent Reoxidation by NAD + Catalyzed by Ferredoxin-NAD + Reductase (Rnf) journal April 2016
Crystal Structure of the 47-kDa Lipoprotein of Treponema pallidum Reveals a Novel Penicillin-binding Protein journal August 2002
Structural Evidence That the 32-Kilodalton Lipoprotein (Tp32) of Treponema pallidum Is an l-Methionine-binding Protein journal October 2004
The PnrA (Tp0319; TmpC) Lipoprotein Represents a New Family of Bacterial Purine Nucleoside Receptor Encoded within an ATP-binding Cassette (ABC)-like Operon in Treponema pallidum journal January 2006
Crystal Structure of the Tp34 (TP0971) Lipoprotein of Treponema pallidum: IMPLICATIONS OF ITS METAL-BOUND STATE AND AFFINITY FOR HUMAN LACTOFERRIN journal December 2006
D-Lactate in Human and Ruminant Metabolism journal July 2005
ETE 3: Reconstruction, Analysis, and Visualization of Phylogenomic Data journal February 2016
Gapped BLAST and PSI-BLAST: a new generation of protein database search programs journal September 1997
PROMALS3D: a tool for multiple protein sequence and structure alignments journal February 2008
Dali server: conservation mapping in 3D journal May 2010
New Algorithms and Methods to Estimate Maximum-Likelihood Phylogenies: Assessing the Performance of PhyML 3.0 journal March 2010
The Microaerophilic Nature of Treponema pallidurn: Enhanced Survival and Incorporation of Tritiated Adenine Under Microaerobic Conditions in the Presence or Absence of Reducing Compounds journal January 1982
The Re-Emergence of Syphilis in the United Kingdom: The New Epidemic Phases journal January 2005
Rising Rates of Syphilis in the Era of Syphilis Elimination journal January 2007
Syphilis Returns to China ??? With a Vengeance journal January 2006
Lipoprotein computational prediction in spirochaetal genomes journal January 2006
Phaser crystallographic software journal July 2007
On the treatment of negative intensity observations journal July 1978
Coot model-building tools for molecular graphics journal November 2004
Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions journal November 2004
MolProbity : all-atom structure validation for macromolecular crystallography journal December 2009
PHENIX: a comprehensive Python-based system for macromolecular structure solution journal January 2010
The CCP4 suite programs for protein crystallography journal September 1994
A novel mode of lactate metabolism in strictly anaerobic bacteria: A novel mode of lactate metabolism in anaerobes journal May 2014
Adaptations of anaerobic archaea to life under extreme energy limitation journal May 2014
Oxidation-reduction potentials of ferredoxin-NADP+ reductase and flavodoxin from Anabaena PCC 7119 and their electrostatic and covalent complexes journal December 1991
Structural basis for stereo-specific catalysis in NAD+-dependent (R)-2-hydroxyglutarate dehydrogenase from Acidaminococcus fermentans: NAD+-dependent (R)-2-hydroxyglutarate dehydrogenase journal December 2004
Bacterial motility: links to the environment and a driving force for microbial physics: Bacterial motility journal January 2006
Complete Genome Sequence of Treponema pallidum, the Syphilis Spirochete journal July 1998
Energetics and Application of Heterotrophy in Acetogenic Bacteria journal May 2016
NADP + -Preferring d-Lactate Dehydrogenase from Sporolactobacillus inulinus journal July 2015
Biological Basis for Syphilis journal January 2006
Lipoproteins of Bacterial Pathogens journal October 2010
Biophysical and Bioinformatic Analyses Implicate the Treponema pallidum Tp34 Lipoprotein (Tp0971) in Transition Metal Homeostasis journal October 2012
Polypeptides of Treponema pallidum: progress toward understanding their structural, functional, and immunologic roles. Treponema Pallidum Polypeptide Research Group. journal January 1993
Sodium Ion Cycle in Bacterial Pathogens: Evidence from Cross-Genome Comparisons journal September 2001
Proposed uniform nomenclature for the proteins and protein domains of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. journal March 1992
The Tp38 (TpMglB-2) Lipoprotein Binds Glucose in a Manner Consistent with Receptor Function in Treponema pallidum journal April 2004
Evidence for Posttranslational Protein Flavinylation in the Syphilis Spirochete Treponema pallidum: Structural and Biochemical Insights from the Catalytic Core of a Periplasmic Flavin-Trafficking Protein journal May 2015
Evidence for an ABC-Type Riboflavin Transporter System in Pathogenic Spirochetes journal February 2013
Long-Term In Vitro Culture of the Syphilis Spirochete Treponema pallidum subsp. pallidum journal June 2018
Motility and Chemotaxis of Spirochetes journal October 1978
Origin and Evolution of the Sodium -Pumping NADH: Ubiquinone Oxidoreductase journal May 2014
The Tp0684 (MglB-2) Lipoprotein of Treponema pallidum: A Glucose-Binding Protein with Divergent Topology journal August 2016
Increase in Incidence of Congenital Syphilis — United States, 2012–2014 journal November 2015
Insights into Domain Closure, Substrate Specificity and Catalysis of D-Lactate Dehydrogenase from Lactobacillus Bulgaricus text January 2002
Insights into Domain Closure, Substrate Specificity and Catalysis of D-Lactate Dehydrogenase from Lactobacillus Bulgaricus text January 2002
a Novel D-lactate Dehydrogenase from Sporolactobacillus sp text January 2015
Flavin-Based Electron Bifurcation, Ferredoxin, Flavodoxin, and Anaerobic Respiration With Protons (Ech) or NAD+ (Rnf) as Electron Acceptors: A Historical Review journal March 2018

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