Transition State Structure and Inhibition of Rv0091, a 5′-Deoxyadenosine/5′-methylthioadenosine Nucleosidase from Mycobacterium tuberculosis

Hilda A. Namanja-Magliano, Christopher F. Stratton, Vern L. Schramm

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

5′-Methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) is a bacterial enzyme that catalyzes the hydrolysis of the N-ribosidic bond in 5′-methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH). MTAN activity has been linked to quorum sensing pathways, polyamine biosynthesis, and adenine salvage. Previously, the coding sequence of Rv0091 was annotated as a putative MTAN in Mycobacterium tuberculosis. Rv0091 was expressed in Escherichia coli, purified to homogeneity, and shown to be a homodimer, consistent with MTANs from other microorganisms. Substrate specificity for Rv0091 gave a preference for 5′-deoxyadenosine relative to MTA or SAH. Intrinsic kinetic isotope effects (KIEs) for the hydrolysis of [1′-3H], [1′-14C], [5′-3H2], [9-15N], and [7-15N]MTA were determined to be 1.207, 1.038, 0.998, 1.021, and 0.998, respectively. A model for the transition state structure of Rv0091 was determined by matching KIE values predicted via quantum chemical calculations to the intrinsic KIEs. The transition state shows a substantial loss of C1′-N9 bond order, well-developed oxocarbenium character of the ribosyl ring, and weak participation of the water nucleophile. Electrostatic potential surface maps for the Rv0091 transition state structure show similarity to DADMe-immucillin transition state analogues. DADMe-immucillin transition state analogues showed strong inhibition of Rv0091, with the most potent inhibitor (5′-hexylthio-DADMe-immucillinA) displaying a Ki value of 87 pM.

Original languageEnglish (US)
Pages (from-to)1669-1676
Number of pages8
JournalACS Chemical Biology
Volume11
Issue number6
DOIs
StatePublished - Jun 17 2016

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adenosylhomocysteine nucleosidase
Mycobacterium tuberculosis
S-Adenosylhomocysteine
Isotopes
Kinetics
Hydrolysis
Salvaging
Quorum Sensing
Nucleophiles
Biosynthesis
Polyamines
Adenine
Substrate Specificity
Static Electricity
Microorganisms
Escherichia coli
Electrostatics
Water
Substrates
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Transition State Structure and Inhibition of Rv0091, a 5′-Deoxyadenosine/5′-methylthioadenosine Nucleosidase from Mycobacterium tuberculosis. / Namanja-Magliano, Hilda A.; Stratton, Christopher F.; Schramm, Vern L.

In: ACS Chemical Biology, Vol. 11, No. 6, 17.06.2016, p. 1669-1676.

Research output: Contribution to journalArticle

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