Fe-S Proteins

Molecular biology and genetic tools to investigate functional redundancy among Fe-S cluster carriers in E. coli.- Study Fe-S cluster proteins in Methanococcus maripaludis, a model archaeal organism.- Genetic approaches to uncover gene products involved in iron-sulfur protein maturation: High throughput genomic screening using transposon-sequencing.- Methods for heterologous overproduction of Fe-S proteins.- Enzymatic and Chemical in vitro Reconstitution of Iron-Sulfur Cluster Proteins.- Expression, isolation, and characterization of vanadium nitrogenase from Azotobacter vinelandii.- Ni-NTA affinity chromatography to characterize protein-protein interactions during Fe-S cluster biogenesis.- tRNA modifications as a readout of S and Fe-S metabolism.- Methods to unravel the roles of ATPases in Fe-S cluster biosynthesis.- Methods to investigate the kinetic profile of cysteine desulfurases.- A fast and ratiometric method for quantification of cysteine-bound persulfides based on alkylation and gel-shift assays.- Circular dichroism to probe the synthesis, transfer, and stability of Fe-S clusters.- Native mass spectrometry of iron-sulfur proteins.- Characterization of Paramagnetic Iron-Sulfur Clusters using Electron Paramagnetic Resonance Spectroscopy.- Characterization of Fe-S clusters in proteins by Mössbauer Spectroscopy.- Biochemical approaches to probe the role of the auxiliary iron-sulfur cluster of lipoyl synthase from Mycobacterium tuberculosis.- Methods to Screen for Radical SAM Enzyme Crystallization Conditions.