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Binding Association and Kinetic Characterization of DesD Reveals High Substrate Specificity and Cooperative Behavior
Author(s) -
Tran Leslie,
Hoffmann Katherine M.
Publication year - 2018
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.2018.32.1_supplement.655.29
Subject(s) - siderophore , isothermal titration calorimetry , biochemistry , enzyme , chemistry , streptomyces coelicolor , enzyme kinetics , kinetics , nonribosomal peptide , substrate (aquarium) , bacteria , biology , stereochemistry , biosynthesis , biophysics , streptomyces , gene , active site , genetics , ecology , physics , quantum mechanics
Siderophores, small iron transporting molecules, are secreted by bacteria when there is an insufficient amount of iron. These molecules have high affinity for ferric ions and often outcompete the host chelators. Siderophores synthetase are becoming increasingly associated with virulence in pathogenic bacteria as they are able to evade the human immune response system. One type of siderophore biosynthesis is through the Nonribosomal peptide synthetase Independent Siderophore (NIS) pathway. This project is interested in utilizing the NIS synthetase family's unique fold and novel chemistry as a drug target for new antibiotics. This project is focused on DesD from Streptomyces coelicolor found mostly in soil. DesD is the last enzyme in the Desferrioxamine E siderophore synthetase pathway. This enzyme is a model for the NIS synthetase family, however, while it has been studied before, there are no good results concerning kinetics. Prior literature has reported a broad substrate specificity but the binding has never been quantified. The preliminary data on binding and kinetics collected from Isothermal Titration Calorimetry (ITC) have revealed high substrate specificity via binding constants, and cooperative kinetic turnover. These baseline studies with be used in future work to quantify inhibitor effectiveness. Support or Funding Information This work was supported by a grant from the National Science Foundation (MCB ‐ 1716986) to K.M.H, and by California Lutheran University. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .