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 applications to protein structure and function and metabolomics

The primary focus of our research is on the structure and function of proteins. We have an NIH-funded project 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), optical 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 from larger macromolecules and their complexes, biological fluids, cell extracts, and natural products. At the BioMagResBank (BMRB,, we maintain the primary depository for biomolecular NMR data. We strive to improve methods for depositing the growing range of data created by biomolecular NMR, and we work with the NMR community in developing better ways of validating the data and making it available in more meaningful ways. In the case of NMR-derived three three-dimensional structures, we partner with the Worldwide Protein Data Bank ( in creating depositions to the Protein Data Bank.

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)