Integrated Approach in Structural Biology and Biophysics

The Stetefeld lab performs a hybrid method approach by combining 3D structural biology techniques with a variety of complementary approaches. In the past, we combined X-ray crystallography with high-resolution NMR techniques underlining the importance of high-resolution data in combination with different time-scale dynamics of extracellular signaling proteins in solution.

At UM, the Stetefeld lab established a new combination of high-resolution structural biology in conjunction with in-depth biophysical characterization of hydrodynamic properties. The combination of different scattering techniques (including dynamic-and static light scattering and small-angle X-ray scattering) in synergy with analytical ultracentrifugation, microscale thermophoresis and interferometric scattering mass spectrometry is ideally suited for the exploration of the dynamic behavior of high-molecular weight protein complexes and for unraveling molecular insights of receptor-ligand formation at near physiological conditions.

It is our goal to implement also low (negative stain) and high-resolution (single particle averaging) cryo-EM techniques into our research program.

In a variety of different projects, we have applied several in silico modeling and structure-based Molecular Dynamics simulations to elucidate highly diverse questions, thereby guiding enhanced diagnosis and therapeutic approaches in the field of extracellular signaling complexes. In collaboration with research groups in the US (NIH), Germany (University of Cologne) and Sweden (Lund University) the Stetefeld laboratory investigated naturally occurring modifications in  human patients in the ACTA1 gene, Collagen XXII and COL12A1 which significantly improved the molecular understanding of these diseases at the molecular level.  Aside, we applied cutting-edge 3D modeling approaches to study cold-adapted O2-transport in extinct mammoth hemoglobin. Most recently, in collaboration with Drs Roshko (Physics) and Viddal (CMU) the sulfur uptake and storage in RHCC-NT have been studied by performing fully atomistic and steered MD simulations in conjunction with umbrella sampling.

The Stetefeld laboratory hosts/ has access to:

  • Advanced eukaryotic cell culture facility and HP-Fermenter system
  • eDNA laboratory (incl. diverse PCR’s/ NGS instrumentation)
  • FPLC-JASCO (FSEC), SEC-MALS-RI and SEC-MALS-QUELS instrumentation
  • Dynamic Light Scattering (DLS) and Circular Dichroism (CD) instrumentation
  • Analytical Ultracentrifuge (AUC)
  • Differential Scanning Calorimetry (vpDSC) and Isothermal Titration Calorimetry (ITC)
  • Microscale Thermophoresis (MST)/ ForteBio Octet K2
  • interferometric Scattering Mass Spectrometry (iSCAMS)
  • Crystallisation facility (incl. several robotics, microscopes and Crystal-Microspectrophotometer)
  • In-house W/SAXS, X-ray Diffractometer and 600MHz NMR instrumentation

In addition, long-term beamtime proposals at the Canadian Light Source (CLS), Advanced Photon Source (APS) and DIAMOND guarantee access to high-brilliance and microfocus beamlines at several synchrotrons and the performance of BIO (SEC)-SAXS experiments.