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Identification of the ligands of TM0486 from Thermotoga maritima by ESI‐TOF mass spectrometry suggests role in a novel thiamin salvage pathway
Author(s) -
Snider Mark J.,
Palanski Brad A,
Rotter Zachary,
Li Xiaopeng
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.923.4
Subject(s) - thermotoga maritima , isothermal titration calorimetry , chemistry , biochemistry , mass spectrometry , electrospray ionization , stereochemistry , computational biology , gene , escherichia coli , biology , chromatography
The structure of the TM0486 gene product from Thermotoga maritima , solved previously without functional annotation, included extra electron density that suggested the presence of ligands that could not be unambiguously identified. We comprehensively screened for ligands released from the purified recombinant protein expressed in E. coli by electrospray ionization time‐of‐flight mass spectrometry and show that thiamin and oxidized forms of thiamin (oxythiamin, desthiothiamin) are bound. Comparative binding studies by isothermal titration calorimetry show that the hydroxymethyl pyrimidine moiety is important, but not sufficient, for binding. These data accord with recent studies suggesting that TM0486 plays a functional role during oxidizing cellular conditions in a novel thiamin salvage pathway. The genomic context of TM0486 includes a gene (TM0484) coding a hypothesized ABC transporter. Current efforts are underway to establish the substrate preference of the ABC transporter and its functional coupling with TM0486. This work was supported by NSF grant #0821110 and by a grant from the Howard Hughes Medical Institute through the Undergraduate Science Education Program to The College of Wooster.