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

Cheng, Yu-Shan; Sacchettini, James C.

doi: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

Journal Article · Fri Feb 05 00:00:00 EST 2016 · Biochemistry

DOI:https://doi.org/10.1021/acs.biochem.5b00993· OSTI ID:1241061

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

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

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.

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

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

Grant/Contract Number:: P01AI095208; A-0015

OSTI ID:: 1241061

Journal Information:: Biochemistry, Vol. 55, Issue 7; ISSN 0006-2960

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 16 works

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

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