Crystallization and preliminary crystallographic analysis of an aminoglycoside kinase from Legionella pneumophila

Abstract

9-Aminoglycoside phosphotransferase type Ia [APH(9)-Ia] is a resistance factor in Legionella pneuemophila, the causative agent of legionnaires' disease. It is responsible for providing intrinsic resistance to the antibiotic spectinomycin. APH(9)-Ia phosphorylates one of the hydroxyl moieties of spectinomycin in an ATP-dependent manner, abolishing the antibiotic properties of this drug. Here, the crystallization and preliminary X-ray studies of this enzyme in two crystal forms is reported. One of the these crystal forms provides diffraction data to a resolution of 1.7 A.

Figure 1

Micrographs of APH(9)-Ia crystals grown in apoenzyme conditions (a), in the presence of ADP and spectinomycin (b) and in the presence of the protein kinase inhibitor CKI-7 (c). The largest crystal dimensions for each were ∼0.8, 0.6 and 0.5 mm, respectively.