faculty

Professor of Chemistry
Ph.D., University of Nijmegen, The Netherlands

NMR Studies of Biomacromolecular Conformation, Dynamics and Interactions in Solution

Phone: (734) 936-3850
Email: zuiderwe@umich.edu 
Fax: 734 747-4865

Research Group

NMR methodolgy has now matured to the point that the systems upto 100 kDa can be studied at atomic level resolution. The proteins are in solution and carry out the conformational changes and dynamic interactions necessary for function while the heteronuclear multi-dimensional NMR experiments are running.

Our group applies all aspects of these methodologies to study Hsp70 protein-folding chaperone proteins in solution. Hsp70Õs mediate trafficking, triaging, folding, unfolding and refolding of other proteins in vivo, from bacteria to man. The groupÕs high-resolution solution structure determinations of Hsp70 domains, as well as studies of atom-resolved dynamics and ligand interactions, have led to the formulation of an allosteric mechanism for these proteins. These mechanisms are being investigated in depth with large constructs (up to 60 kDa) that have become accessible for TROSY NMR studies by using 800 MHz equipment. We discover that these active allosteric constructs in solution change their conformation in upon ligand binding. We are also investigating complexes of chaperone and co-chaperone proteins in different allosteric states

Dynamics (local motion) is an essential component of biological functioning. Without motion, proteins cannot accommodate ligands, carry out chemistry, be allosterically active or be thermally stable. The group is working on the fundamental problem of the modeling of local motions. In several small proteins, the group has found evidence for correlated motions, for conformational change dynamics in enzyme active sites and for large changes in entropy contained in the dynamics of the protein backbone upon ligand binding.

The laboratory has dedicated 500 and 600 MHz NMR spectrometers, and we are main users of a another 600 MHz and a 800 MHz NMR spectrometer. The state-of-the-art instruments are equipped with four radio-frequency channels and (cryogenic) triple-resonance gradient probes for the execution of the most modern multi-dimensional NMR experiments. A large Silicon Graphics / P.C. computer cluster is available for data processing, spectral interpretation and structure display. We have two wet-labs for protein mutagenesis, expression, purification and sample preparation.

Our group is housed in the Chemestry builiding on Central Campus. Together with two other biological NMR groups in that builiding (Al-Hashimi and Ramamoorthy) we form a vibrant NMR research community of about twenty-five students and post-docs, with combined group meetings, parties and specialized seminars.

REPRESENTATIVE PUBLICATIONS

1. T Wang, S Cai, ERP Zuiderweg. "Temperature dependence of anisotropic protein backbone dynamics" J. Am. Chem. Soc. 2003, 125, 8639.
2. P Khandelwal, K Keliikuli, CL Smith, MA Saper, ERP Zuiderweg. "Solution structure and phosphopeptide binding to the N-terminal domain of Yersinia YopH, Comparison with a Crystal Structure" Biochemistry, 2002, 41, 11425.
3. SY Stevens, S Sanker, C Kent, ERP Zuiderweg. "Delineation of the allosteric mechanism for a cytidylyltransferase exhibiting negative cooperativity" Nature Structural Biology 2001, 8, 947.
4. M Pellecchia, SY Stevens, CW Vander Kooi, DH Montgomery, EH Feng, LM Gierasch ERP Zuiderweg. "Structural insights into substrate binding by the molecular chaperone DnaK" Nature Structural Biology, 2000, 7, 298
5. RC Morshauser, W Hu, H Wang, Y Pang, GC Flynn, ERP Zuiderweg. "High resolution solution structure of the 18 kda substrate binding domain of the mammalian chaperone protein hsc" J. Mol. Biol. 1999, 289, 1387