Faculty Summaries
Heinrich Roder, PhD
Heinrich Roder, PhD
Professor
  • Adjunct Professor, University of Pennsylvania
  • Professor, Temple University
  • Director, Spectroscopy Support Facility
Heinrich.Roder@fccc.edu
Office Phone: 215-728-3123
Fax: 215-728-3574
Office: R414
Lab: R407/408
Protein Folding, Structure and Function

A central theme of our research concerns the early stages of protein folding, which are critical for understanding how the native structure of a protein, and ultimately its function, are encoded in the amino acid sequence. The insight gained not only provides a basis for protein structure prediction and de novo design, but also contributes to our mechanistic understanding and treatment of a wide range of diseases that involve aggregation of denatured or misfolded proteins. The stability and folding dynamics of proteins also has major implications with respect to understanding of the physiological consequences of mutations, in vivo folding and other cellular processes, such as trafficking and degradation. We study the dynamics of protein folding on a microsecond time scale by coupling advanced mixing techniques with various detection methods, including fluorescence and H/D exchange labeling experiments with NMR detection. In combination with protein engineering, these approaches have provided detailed insight into the folding mechanisms of a diverse set of proteins [reviewed in Roder et al., 2006]. These approaches are currently being used to elucidate the sequence determinants for folding initiation and propagation of apomyoglobin, a prototypic alpha-helical protein [Xu et al., 2012].Quenched-flow H/D exchange measurements, using a novel microfludic mixing device, allow us to detect the formation of individual hydrogen bonds in early folding intermediates populated within 100 μs of refolding. Recent results indicate that the initial compaction of cytochrome c involves specific formation of secondary and tertiary structure rather than a non-specific hydrophobic collapse [Fazelinia et al., 2014].

Our group also investigates the structure, dynamics and molecular interactions of various proteins of biomedical interest in solution by using NMR spectroscopy and other biophysical methods. For example, we found that zymogen activation of factor XI (triggered by cleavage of a scissile bond) is accompanied by a major change in overall shape of this large multi-domain protein [Samuel et al., 2007]. Other studies focus on the signaling adaptor protein Na+/H+ exchanger regulatory factor 1 (NHERF1), which plays a central role in cellular pathways relating receptors and ion channels with the actin cytoskeleton. NHERF1 contains two globular domains belonging to the PDZ domain family and a C-terminal ezrin binding (EB) motif, as well as two long segments predicted to be intrinsically disordered. Using NMR, circular dichroism and fluorescence methods, we have shown that NHERF1 can adopt two alternative closed conformations in which the second PDZ domain engages the C-terminal region via eith non-specific interactions with the disordered linker or specific contacts with the EB motif [Cheng et al., 2009]. The initially unstructured EB region becomes helical when bound to the second PDZ domain. Together with equilibrium unfolding and ligand binding data, the findings indicate that the activity of NHERF as a signaling adaptor is regulated by a subtle balance between competing intra- and intermolecular domain-domain interactions.

Description of research projects
Selected Publications
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  1. Fazelinia, H., Xu, M., Cheng, H., Roder, H. Ultrafast hydrogen exchange reveals specific structural events during the initial stages of folding of cytochrome c. J. Am. Chem. Soc., in press. JACS PubMed
  2. Montalvo, G. L., Gai, F., Roder, H., Degrado, W. F. Slow Folding-Unfolding Kinetics of an Octameric beta-Peptide Bundle. ACS Chem Biol. 2014; 9:276-81. PubMed
  3. Mizukami, T., Xu, M., Cheng, H., Roder, H., Maki, K. Non-uniform chain collapse during early stages of staphylococcal nuclease folding by fluorescence resonance energy transfer and ultrarapid mixing methods. Protein Science 2013; 22:1336-48. PubMed
  4. Moroz, O.V., Moroz, Y. S., Wu, Y., Olsen, A. B., Cheng, H., Mack, K. L., McLaughlin, J. M., Raymond, E. A., Zhezherya, K., Roder, H, Korendovych, I. V. A Single Mutation in a Regulatory Protein Produces Evolvable Allosterically Regulated Catalyst of Nonnatural Reaction. Angew. Chem. Int. Ed. Engl. 2013; 52:6246-6249. PubMed
  5. Xu M, Beresneva O, Rosario R, Roder H. Microsecond Folding Dynamics of Apomyoglobin at Acidic pH. J Phys Chem B. 2012; 116(23):7014-25. PubMed
  6. Korendovych I V, Kulp DW, Wu Y, Cheng H, Roder H, DeGrado WF. Design of a switchable eliminase. Proc Natl Acad Sci U S A. 2011; 108:6823-7. PubMed
  7. Chen KC, Xu M, Wedemeyer WJ, Roder H. Microsecond unfolding kinetics of sheep prion protein reveals an intermediate that correlates with susceptibility to classical scrapie. Biophys J. 2011;101:1221-30. PubMed
  8. Katz, R. A., Merkel, G., Andrake, M. D., Roder, H. & Skalka, A. M. Retroviral integrases promote fraying of viral DNA ends. J. Biol. Chem.2011; 286:25710-8 PubMed
  9. Lau WL, DeGrado WF, Roder H. The effects of pKa tuning on the thermodynamics and kinetics of folding: Design of a solvent-shielded carboxylate pair at the “a”position of a coiled coil. Biophys J. 2010; 99:2299-2308. PubMed
  10. Alves C, Cheng H, Roder H, Taylor J. Intrinsic disorder and oligomerization of the hepatitis delta virus antigen. Virology. 2010; 407:333-340. PubMed
  11. Cheng H, Li J, Fazlieva R, Dai Z, Bu Z, Roder H. Autoinhibitory interactions between the PDZ2 and C-terminal domains in the scaffolding protein NHERF1. Structure. 2009;17(5):660-669. PubMed
  12. Latypov RF, Maki K, Cheng H, Luck SD, Roder H. Folding mechanism of reduced Cytochrome c: equilibrium and kinetic properties in the presence of carbon monoxide. J Mol Biol. 2008;383(2):437-53. PubMed
  13. Maki K, Cheng H, Dolgikh DA, Roder H. Folding kinetics of staphylococcal nuclease studied by tryptophan engineering and rapid mixing methods. J Mol Biol. 2007;368(1):244-55. PubMed
  14. Samuel D, Cheng H, Riley PW, Canutescu AA, Nagaswami C, Weisel JW, Bu Z, Walsh PN, Roder H. Solution structure of the A4 domain of factor XI sheds light on the mechanism of zymogen activation. Proc Natl Acad Sci USA. 2007;104(40):15693-8. PubMed
  15. Apetri AC, Maki K, Roder H, Surewicz WK. Early intermediate in human prion protein folding as evidenced by ultrarapid mixing experiments. J Am Chem Soc. 2006;128(35):11673-8. PubMed
  16. Latypov RF, Cheng H, Roder NA, Zhang J, Roder H. Structural characterization of an equilibrium unfolding intermediate in cytochrome c. J Mol Biol. 2006;357(3):1009-25. PubMed
  17. Roder H, Maki K, Cheng H. Early events in protein folding explored by rapid mixing methods. Chem Rev. 2006;106(5):1836-61. PubMed
All publications