John L. Markley

Photo of John L. Markley
Steenbock Professor of Biomolecular Structure (also Director of NMRFAM and BMRB)
B.A., Carleton College
Ph.D., Harvard University
NIH Postdoctoral Fellow, University of California, Berkeley
Phone: (608) 263-9349

NMR spectroscopy and its biological applications; structure function relationships in proteins; stable-isotope-assisted multinuclear magnetic resonance spectroscopy; processing and analysis of multi-dimensional NMR data; structural genomics; metabolomics

The primary focus of our research is on the structure and function of proteins. We have an NIH-funded project on mitochondrial proteins (, where we are identifying protein-protein interactions and elucidating functional properties of proteins. Much of our recent work has been on the protein machinery involved in the biosynthesis and delivery of iron-sulfur clusters. We rely on NMR spectroscopy as a major approach to studying protein structure and dynamics and supplement this with information from small angle x-ray scattering (SAXS), differential scanning calorimetry (DSC), oprical spectroscopy, and other biophysical approaches. At the National Magnetic Resonance Facility at Madison (NMRFAM,, we develop technology as driven by collaborative investigations on improved methods for collecting and analyzing NMR data, larger macromolecules and their complexes, and metabolomics and natural products. At the Center for Eukaryotic Structural Genomics (CESG,, we develop and apply technology for protein production and labeling with stable isotopes for NMR investigations and Se-Met for x-ray crystallography. At the BioMagResBank (BMRB,, we work to improve ways of annotating and archiving NMR data and associated information along with three-dimensional structures as part of the Worldwide Protein Data Bank. At the Accelerated Renewable Resources Consortium (AREC,, we are carrying out a research and demonstration project on the conversion of dairy manure into mulch, biofuels, fertilizer pellets, liquid fertilizer, and other commodities.

image of co chaperone proteins HscB and HscA

Information from NMR spectroscopy on the nucleotide-dependent interaction between the co-chaperone protein (HscB) shown above and the chaperone protein HscA. We used the T212V mutant of HscA, which lacks ATPase activity. NMR signal perturbation profiles of [U-15N]-HscB with HscA(T212V) mapped onto the structure of HscB (PDB 1FPO). Color code: (black) residues with no signal (Pro), unassigned residues, or residues whose signals could not be followed upon addition of HscA(T212V); (gray) residues whose signals were minimally affected (DdNH < 0.01 ppm) by addition of HscA(T212V); (blue) residues with DdNH > 0.01 ppm; and (red) residues whose signals broadened beyond detection. Only the surface of the structure is shown to better represent the putative HscA binding interface.

From: J. Am. Chem. Soc. 136, 11586-11589 (2014)