Techniques
The Manley lab uses a variety of bioinformatic, biochemical, biophysical, structural biology, analytical, and spectroscopic techniques to answer questions about natural product biosynthesis and enzymology.
Bioinformatics
Bacterial genomic information reveals a plethora of untapped genes with the potential to encode exciting metalloenzyme chemistry. The Manley lab uses genome mining strategies to comb through genomic information and identify yet uncharacterized metalloenzymes, focusing on those associated with the biosynthesis of novel natural products.
Enzymology
Biochemical assays are used to study the reactions performed by enzymes. We can monitor reaction progress using techniques such as UV-visible spectroscopy, liquid chromatography, and mass spectrometry. To monitor rapid reactions, the Manley lab uses stopped-flow spectroscopy which can track reactions on a millisecond time-scale.
Natural Product Characterization
For enzymes involved in the biosynthesis of novel natural products, the structure of the product must be elucidated. We can use structural tools such as mass spectrometry and nuclear magnetic resonance spectroscopy to determine the structures of novel compounds.
Structural Biology
To gain insight into the structure-function relationships of enzymes, we rely on structural biology techniques such as X-ray crystallography.
Spectroscopy
The metal cofactors of the enzymes in the Manley lab make them amenable to investigation by spectroscopic techniques such as electron paramagnetic resonance (EPR) spectroscopy, Mössbauer spectroscopy, and more. These techniques will shed light on the nature of the metal center and its role during the enzyme's catalytic cycle.