Uncovering Zn²⁺ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d-2-hydroxyglutarate dehydrogenase.

ABSTRACT: Pseudomonas aeruginosa couples the oxidation of d-2-hydroxyglutarate (D2HG) to l-serine biosynthesis for survival, using d-2-hydroxyglutarate dehydrogenase from P. aeruginosa (PaD2HGDH). Knockout of PaD2HGDH impedes P. aeruginosa growth, making PaD2HGDH a potential target for therapeutics. Previous studies showed that the enzyme's activity increased with Zn²⁺, Co²⁺, or Mn²⁺ but did not establish the enzyme's metal composition and whether the metal is an activator or a required cofactor for the enzyme, which we addressed in this study. Comparable to the human enzyme, PaD2HGDH showed only 15% flavin reduction with D2HG or d-malate. Upon purifying PaD2HGDH with 1 mM Zn²⁺, the Zn²⁺:protein stoichiometry was 2:1, yielding an enzyme with ∼40

SUBMITTER: Quaye JA 

PROVIDER: S-EPMC10025160 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature