Structural and Functional Characterization of Malate Synthase G from Opportunistic Pathogen Pseudomonas aeruginosa.
Alyssa C McVeyPrasanthi MedarametlaXavier CheeSean BartlettAntti PosoDavid R SpringTaufiq RahmanMartin WelchPublished in: Biochemistry (2017)
Pseudomonas aeruginosa is an opportunistic human pathogen recognized as a critical threat by the World Health Organization because of the dwindling number of effective therapies available to treat infections. Over the past decade, it has become apparent that the glyoxylate shunt plays a vital role in sustaining P. aeruginosa during infection scenarios. The glyoxylate shunt comprises two enzymes: isocitrate lyase and malate synthase isoform G. Inactivation of these enzymes has been reported to abolish the ability of P. aeruginosa to establish infection in a mammalian model system, yet we still lack the structural information to support drug design efforts. In this work, we describe the first X-ray crystal structure of P. aeruginosa malate synthase G in the apo form at 1.62 Å resolution. The enzyme is a monomer composed of four domains and is highly conserved with homologues found in other clinically relevant microorganisms. It is also dependent on Mg2+ for catalysis. Metal ion binding led to a change in the intrinsic fluorescence of the protein, allowing us to quantitate its affinity for Mg2+. We also identified putative drug binding sites in malate synthase G using computational analysis and, because of the high resolution of the experimental data, were further able to characterize its hydration properties. Our data reveal two promising binding pockets in malate synthase G that may be exploited for drug design.
Keyphrases
- pseudomonas aeruginosa
- high resolution
- cystic fibrosis
- electronic health record
- biofilm formation
- endothelial cells
- big data
- candida albicans
- acinetobacter baumannii
- climate change
- pulmonary artery
- transcription factor
- healthcare
- genome wide
- dna binding
- magnetic resonance imaging
- escherichia coli
- gene expression
- data analysis
- computed tomography
- pulmonary hypertension
- magnetic resonance
- health information
- artificial intelligence
- deep learning
- social media
- high speed