Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance

doi:https://doi.org/10.1021/acs.biochem.5b00993

Title: Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance

Abstract

Mycobacterium tuberculosis ( Mtb) Rv2671 is annotated as a 5-amino-6-ribitylamino-2,4(1 H,3 H)-pyrimidinedione 5'-phosphate (AROPP) reductase (RibD) in the riboflavin biosynthetic pathway. Recently, a strain of Mtb with a mutation in the 5' untranslated region of Rv2671, which resulted in its overexpression, was found to be resistant to dihydrofolate reductase (DHFR) inhibitors including the anti- Mtb drug para-aminosalicylic acid (PAS). In this study, a biochemical analysis of Rv2671 showed that it was able to catalyze the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), which explained why the overexpression of Rv2671 was sufficient to confer PAS resistance. We solved the structure of Rv2671 in complex with the NADP ⁺ and tetrahydrofolate (THF), which revealed the structural basis for the DHFR activity. Furthermore, the structures of Rv2671 complexed with two DHFR inhibitors, trimethoprim and trimetrexate, provided additional details of the substrate binding pocket and elucidated the differences between their inhibitory activities. Finally, Rv2671 was unable to catalyze the reduction of AROPP, which indicated that Rv2671 and its closely related orthologues are not involved in riboflavin biosynthesis.

Authors:

Cheng, Yu-Shan ^[1]; Sacchettini, James C. ^[1]

Texas A & M Univ., College Station, TX (United States)

Publication Date:: 2016-02-05

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Org.:: National Institutes of Health (NIH); R.J. Wolfe-Welch Foundation

OSTI Identifier:: 1241061

Grant/Contract Number:: P01AI095208; A-0015

Resource Type:: Journal Article: Accepted Manuscript

Journal Name:: Biochemistry

Additional Journal Information:: Journal Volume: 55; Journal Issue: 7; Journal ID: ISSN 0006-2960

Publisher:: American Chemical Society (ACS)

Country of Publication:: United States

Language:: ENGLISH

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Peptides and proteins; Pyrimidine; Chemical structure; Inhibitors; Noncovalent interactions

Citation Formats


                    Cheng, Yu-Shan, and Sacchettini, James C. Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance.  United States: N. p., 2016. 
        Web.  doi:10.1021/acs.biochem.5b00993.

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                    Cheng, Yu-Shan, & Sacchettini, James C. Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance.  United States.  https://doi.org/10.1021/acs.biochem.5b00993

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                    Cheng, Yu-Shan, and Sacchettini, James C. Fri .  
        "Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance".  United States.  https://doi.org/10.1021/acs.biochem.5b00993.  https://www.osti.gov/servlets/purl/1241061.

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@article{osti_1241061,

  title        = {Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance},

  author       = {Cheng, Yu-Shan and Sacchettini, James C.},

  abstractNote = {Mycobacterium tuberculosis (Mtb) Rv2671 is annotated as a 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione 5'-phosphate (AROPP) reductase (RibD) in the riboflavin biosynthetic pathway. Recently, a strain of Mtb with a mutation in the 5' untranslated region of Rv2671, which resulted in its overexpression, was found to be resistant to dihydrofolate reductase (DHFR) inhibitors including the anti-Mtb drug para-aminosalicylic acid (PAS). In this study, a biochemical analysis of Rv2671 showed that it was able to catalyze the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), which explained why the overexpression of Rv2671 was sufficient to confer PAS resistance. We solved the structure of Rv2671 in complex with the NADP+ and tetrahydrofolate (THF), which revealed the structural basis for the DHFR activity. Furthermore, the structures of Rv2671 complexed with two DHFR inhibitors, trimethoprim and trimetrexate, provided additional details of the substrate binding pocket and elucidated the differences between their inhibitory activities. Finally, Rv2671 was unable to catalyze the reduction of AROPP, which indicated that Rv2671 and its closely related orthologues are not involved in riboflavin biosynthesis.},

  doi          = {10.1021/acs.biochem.5b00993},

  url          = {https://www.osti.gov/biblio/1241061},
  journal      = {Biochemistry},

  issn         = {0006-2960},

  number       = 7,

  volume       = 55,

  place        = {United States},

  year         = {2016},

  month        = {2}

}

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https://doi.org/10.1021/acs.biochem.5b00993

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Title: Structural Insights into Mycobacterium tuberculosis Rv2671 Protein as a Dihydrofolate Reductase Functional Analogue Contributing to para -Aminosalicylic Acid Resistance

Abstract

Citation Formats

Identification of potent chromone embedded [1,2,3]-triazoles as novel anti-tubercular agents journal, April 2018

Identification of potent chromone embedded [1,2,3]-triazoles as novel anti-tubercular agents
journal, April 2018