Characterization of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi: Antibiotic targets for Lyme disease

Kenneth A. Cornell, Reece J. Knippel, Gerald R. Cortright, Meghan Fonken, Christian Guerrero, Amy R. Hall, Kristen A. Mitchell, John H. Thurston, Patrick Erstad, Aoxiang Tao, Dong Xu, Nikhat Parveen

Research output: Contribution to journalArticle

Abstract

Background: Borrelia burgdorferi causes Lyme disease, the most common tick-borne illness in the United States. The Center for Disease Control and Prevention estimates that the occurrence of Lyme disease in the U.S. has now reached approximately 300,000 cases annually. Early stage Borrelia burgdorferi infections are generally treatable with oral antibiotics, but late stage disease is more difficult to treat and more likely to lead to post-treatment Lyme disease syndrome. Methods: Here we examine three unique 5′-methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidases (MTNs or MTANs, EC 3.2.2.9) responsible for salvage of adenine and methionine in B. burgdorferi and explore their potential as antibiotic targets to treat Lyme disease. Recombinant Borrelia MTNs were expressed and purified from E. coli. The enzymes were extensively characterized for activity, specificity, and inhibition using a UV spectrophotometric assay. In vitro antibiotic activities of MTN inhibitors were assessed using a bioluminescent BacTiter-Glo™ assay. Results: The three Borrelia MTNs showed unique activities against the native substrates MTA, SAH, and 5′-deoxyadenosine. Analysis of substrate analogs revealed that specific activity rapidly dropped as the length of the 5′-alkylthio substitution increased. Non-hydrolysable nucleoside transition state analogs demonstrated sub-nanomolar enzyme inhibition constants. Lastly, two late stage transition state analogs exerted in vitro IC50 values of 0.3–0.4 μg/mL against cultured B. burgdorferi cells. Conclusion: B. burgdorferi is unusual in that it expresses three distinct MTNs (cytoplasmic, membrane bound, and secreted) that are effectively inactivated by nucleoside analogs. General significance: The Borrelia MTNs appear to be promising targets for developing new antibiotics to treat Lyme disease.

Original languageEnglish (US)
Article number129455
JournalBiochimica et Biophysica Acta - General Subjects
Volume1864
Issue number1
DOIs
StatePublished - Jan 2020

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adenosylhomocysteine nucleosidase
Borrelia burgdorferi
Lyme Disease
Borrelia
Anti-Bacterial Agents
Nucleosides
Pemetrexed
Borrelia Infections
Assays
Disease control
Enzyme inhibition
Salvaging
Ticks
Adenine
Enzymes
Centers for Disease Control and Prevention (U.S.)
Methionine
Inhibitory Concentration 50
Substrates
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Keywords

  • Borrelia
  • Lyme disease
  • MTA/SAH Nucleosidases
  • MTAN
  • MTN
  • Transition state analog

Cite this

Cornell, Kenneth A. ; Knippel, Reece J. ; Cortright, Gerald R. ; Fonken, Meghan ; Guerrero, Christian ; Hall, Amy R. ; Mitchell, Kristen A. ; Thurston, John H. ; Erstad, Patrick ; Tao, Aoxiang ; Xu, Dong ; Parveen, Nikhat. / Characterization of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi : Antibiotic targets for Lyme disease. In: Biochimica et Biophysica Acta - General Subjects. 2020 ; Vol. 1864, No. 1.
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abstract = "Background: Borrelia burgdorferi causes Lyme disease, the most common tick-borne illness in the United States. The Center for Disease Control and Prevention estimates that the occurrence of Lyme disease in the U.S. has now reached approximately 300,000 cases annually. Early stage Borrelia burgdorferi infections are generally treatable with oral antibiotics, but late stage disease is more difficult to treat and more likely to lead to post-treatment Lyme disease syndrome. Methods: Here we examine three unique 5′-methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidases (MTNs or MTANs, EC 3.2.2.9) responsible for salvage of adenine and methionine in B. burgdorferi and explore their potential as antibiotic targets to treat Lyme disease. Recombinant Borrelia MTNs were expressed and purified from E. coli. The enzymes were extensively characterized for activity, specificity, and inhibition using a UV spectrophotometric assay. In vitro antibiotic activities of MTN inhibitors were assessed using a bioluminescent BacTiter-Glo™ assay. Results: The three Borrelia MTNs showed unique activities against the native substrates MTA, SAH, and 5′-deoxyadenosine. Analysis of substrate analogs revealed that specific activity rapidly dropped as the length of the 5′-alkylthio substitution increased. Non-hydrolysable nucleoside transition state analogs demonstrated sub-nanomolar enzyme inhibition constants. Lastly, two late stage transition state analogs exerted in vitro IC50 values of 0.3–0.4 μg/mL against cultured B. burgdorferi cells. Conclusion: B. burgdorferi is unusual in that it expresses three distinct MTNs (cytoplasmic, membrane bound, and secreted) that are effectively inactivated by nucleoside analogs. General significance: The Borrelia MTNs appear to be promising targets for developing new antibiotics to treat Lyme disease.",
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author = "Cornell, {Kenneth A.} and Knippel, {Reece J.} and Cortright, {Gerald R.} and Meghan Fonken and Christian Guerrero and Hall, {Amy R.} and Mitchell, {Kristen A.} and Thurston, {John H.} and Patrick Erstad and Aoxiang Tao and Dong Xu and Nikhat Parveen",
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Cornell, KA, Knippel, RJ, Cortright, GR, Fonken, M, Guerrero, C, Hall, AR, Mitchell, KA, Thurston, JH, Erstad, P, Tao, A, Xu, D & Parveen, N 2020, 'Characterization of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi: Antibiotic targets for Lyme disease', Biochimica et Biophysica Acta - General Subjects, vol. 1864, no. 1, 129455. https://doi.org/10.1016/j.bbagen.2019.129455

Characterization of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi : Antibiotic targets for Lyme disease. / Cornell, Kenneth A.; Knippel, Reece J.; Cortright, Gerald R.; Fonken, Meghan; Guerrero, Christian; Hall, Amy R.; Mitchell, Kristen A.; Thurston, John H.; Erstad, Patrick; Tao, Aoxiang; Xu, Dong; Parveen, Nikhat.

In: Biochimica et Biophysica Acta - General Subjects, Vol. 1864, No. 1, 129455, 01.2020.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidases from Borrelia burgdorferi

T2 - Antibiotic targets for Lyme disease

AU - Cornell, Kenneth A.

AU - Knippel, Reece J.

AU - Cortright, Gerald R.

AU - Fonken, Meghan

AU - Guerrero, Christian

AU - Hall, Amy R.

AU - Mitchell, Kristen A.

AU - Thurston, John H.

AU - Erstad, Patrick

AU - Tao, Aoxiang

AU - Xu, Dong

AU - Parveen, Nikhat

PY - 2020/1

Y1 - 2020/1

N2 - Background: Borrelia burgdorferi causes Lyme disease, the most common tick-borne illness in the United States. The Center for Disease Control and Prevention estimates that the occurrence of Lyme disease in the U.S. has now reached approximately 300,000 cases annually. Early stage Borrelia burgdorferi infections are generally treatable with oral antibiotics, but late stage disease is more difficult to treat and more likely to lead to post-treatment Lyme disease syndrome. Methods: Here we examine three unique 5′-methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidases (MTNs or MTANs, EC 3.2.2.9) responsible for salvage of adenine and methionine in B. burgdorferi and explore their potential as antibiotic targets to treat Lyme disease. Recombinant Borrelia MTNs were expressed and purified from E. coli. The enzymes were extensively characterized for activity, specificity, and inhibition using a UV spectrophotometric assay. In vitro antibiotic activities of MTN inhibitors were assessed using a bioluminescent BacTiter-Glo™ assay. Results: The three Borrelia MTNs showed unique activities against the native substrates MTA, SAH, and 5′-deoxyadenosine. Analysis of substrate analogs revealed that specific activity rapidly dropped as the length of the 5′-alkylthio substitution increased. Non-hydrolysable nucleoside transition state analogs demonstrated sub-nanomolar enzyme inhibition constants. Lastly, two late stage transition state analogs exerted in vitro IC50 values of 0.3–0.4 μg/mL against cultured B. burgdorferi cells. Conclusion: B. burgdorferi is unusual in that it expresses three distinct MTNs (cytoplasmic, membrane bound, and secreted) that are effectively inactivated by nucleoside analogs. General significance: The Borrelia MTNs appear to be promising targets for developing new antibiotics to treat Lyme disease.

AB - Background: Borrelia burgdorferi causes Lyme disease, the most common tick-borne illness in the United States. The Center for Disease Control and Prevention estimates that the occurrence of Lyme disease in the U.S. has now reached approximately 300,000 cases annually. Early stage Borrelia burgdorferi infections are generally treatable with oral antibiotics, but late stage disease is more difficult to treat and more likely to lead to post-treatment Lyme disease syndrome. Methods: Here we examine three unique 5′-methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidases (MTNs or MTANs, EC 3.2.2.9) responsible for salvage of adenine and methionine in B. burgdorferi and explore their potential as antibiotic targets to treat Lyme disease. Recombinant Borrelia MTNs were expressed and purified from E. coli. The enzymes were extensively characterized for activity, specificity, and inhibition using a UV spectrophotometric assay. In vitro antibiotic activities of MTN inhibitors were assessed using a bioluminescent BacTiter-Glo™ assay. Results: The three Borrelia MTNs showed unique activities against the native substrates MTA, SAH, and 5′-deoxyadenosine. Analysis of substrate analogs revealed that specific activity rapidly dropped as the length of the 5′-alkylthio substitution increased. Non-hydrolysable nucleoside transition state analogs demonstrated sub-nanomolar enzyme inhibition constants. Lastly, two late stage transition state analogs exerted in vitro IC50 values of 0.3–0.4 μg/mL against cultured B. burgdorferi cells. Conclusion: B. burgdorferi is unusual in that it expresses three distinct MTNs (cytoplasmic, membrane bound, and secreted) that are effectively inactivated by nucleoside analogs. General significance: The Borrelia MTNs appear to be promising targets for developing new antibiotics to treat Lyme disease.

KW - Borrelia

KW - Lyme disease

KW - MTA/SAH Nucleosidases

KW - MTAN

KW - MTN

KW - Transition state analog

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