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Diadenosine Polyphosphatases of the NUDIX Hydrolase Superfamily in M. tuberculosis
Author(s) -
DiCola Andrew Michael,
Knowles Alexander,
O'Handley Suzanne
Publication year - 2017
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.31.1_supplement.939.6
Subject(s) - enzyme , tuberculosis , hydrolase , divalent , biology , virulence , chemistry , biochemistry , gene , medicine , organic chemistry , pathology
M. tuberculosis contains 11 potential Nudix hydrolases, and we are characterizing these enzymes as potential novel antibiotic targets. The diadenosine polyphosphatases (Ap n Aases)/mRNA decapping enzymes are a family of enzymes within the Nudix hydrolase superfamily. In M. tuberculosis there is the primary Nudix Ap n Aase and the secondary Nudix Ap n Aase. The diadenosine polyphosphatases from Legionella pneumophilia and Bartonella bacilliformis have been found to be important in each pathogen's ability to invade its host cells. It is of interest to know whether these enzymes act in the same way in M. tuberculosis. If they are all found to be involved in invasiveness and thus in virulence, then these enzymes could be novel antibiotic targets. We have cloned and overexpressed each protein and have subcloned each into a HisTag vector to optimize purification. We have purified the secondary Nudix Ap n Aase from M. tuberculosis so that we can complete its characterization. Thus far we have determined its substrate specificity (Ap 4 A is the best substrate), Ap n Aase activity, optimal pH (7.8), and divalent metal ion requirements (1 mM Mn 2+ yields optimal activity, however Mg 2+ , Co 2+ , and Zn 2+ yield activity as well). Support or Funding Information This research has been supported by an NIH AREA grant and RIT COS and honors summer undergraduate research fellowships.