RIS Members - John J. Osterhout

Folding and Design Laboratory

Members

John Osterhout
Sunita Kulkarni

Research

alpha-t-alpha
Peptide Dissection
HX

Group Alumni

Youcef Fezoui
Marcela Oslin
Tanya Knubovets
Diane Schaak
Wujing Xian

John J. Osterhout

Protein Folding and Design Laboratory

Contact Information

Research Associate Professor
Institute for Biomedical Science and Biotechnology
BSW 362
P.O. Box 210088
University of Arizona
Tucson, Arizona 85721

Telephone: 520-626-1311
E-mail: johno@email.arizona.edu

Research Interests

Recent fast kinetic experiments show that proteins can fold rapidly (on the millisecond time scale) in the absence of kinetic blocks such as proline isomerization or disulfide formation. Both the old view of protein folding (intermediates/pathways) and the new view (ensembles/energy landscapes) provide suggestions about how proteins might dodge the Levinthal paradox and manage to attain their native conformations on time scales less than eons. However, no matter which view of protein folding one prefers, a correlation between structure, stability, and the manner in which short and long range interactions conspire to direct folding along a pathway or funnel toward the native state is still desirable. This is a difficult problem to attack experimentally because the interesting intermediates are only transiently populated in kinetic experiments and fractionally populated in equilibrium experiments.

Our approach to this problem is to use peptides as model systems. Our primary interest is the study of folding at the level of secondary structure association. To this end, a 38-residue peptide, alpha-t-alpha was designed de novo to form a helical hairpin in solution. The success of this design was confirmed by NMR structure determination. Current research is directed toward understanding the balance of forces contributing to the stability of the molecule and toward measuring the kinetics of helix association.

Publications

Alpha-t-alpha Publications

Fezoui, Y, Hartley, D. M., Walsh, D. M., Selkoe, D. J., Osterhout, J. J. and Teplow, D. B. (2000) A de novo designed helix-turn-helix peptide forms nontoxic amyloid fibrils. Nature Structural Biology 7, 1095-1099

Fezoui, Y., Braswell, E. H., Xian, W. and Osterhout, J. J. (1999) Dissection of the de Novo Designed Peptide alpha-t-alpha: Stability and Properties of the Intact Molecule and Its Constituent Helices. Biochemistry 38, 2796-2804

Fezoui, Y., Connolly, P. J. and Osterhout, J. J. (1997) Solution Structure of alpha-t-alpha, a Helical Hairpin Peptide of De Novo Design. Protein Science 6, 1869-1877

Fezoui, Y., Weaver, D. L. and Osterhout, J. J. (1995). Strategies and Rationales for the De Novo Design of a Helical Hairpin Peptide. Protein Science 4, 286-295

Fezoui, Y., Weaver, D. L. and Osterhout, J. J. (1994). De Novo Design and Structural Characterization of an alpha-Helical Hairpin Peptide - A Model System for the Study of Protein Folding Intermediates. Proc. Natl. Acad. Sci. USA 91, 3675-3679

Protein Folding Publications

Knubovets, T., Osterhout, J. J., Connolly, P. J. and Klibanov, A. M. (1999). Structure, thermostability, and conformational flexibility of hen egg-white lysozyme dissolved in glycerol. Proc. Natl. Acad. Sci. USA 96, 1262-1267

Knubovets, T., Osterhout, J. J. and Klibanov, A. M. (1999). Structure of Lysozyme Dissolved in Neat Organic Solvents as Assessed by NMR and CD Spectroscopies. Biotechnology and Bioengineering 63, 242-248

Choe, S. E., Matsudaira, P. T., Osterhout, J., Wagner, G., and Shakhnovich, E. I., (1998). Folding Kinetics of Villin 14T, a Protein Domain with a Central beta-Sheet and Two Hydrophobic Cores. Biochemistry 37, 14508-14518

Dyson, H. J., Bolinger, L., Feher, V. A. Osterhout, J. J. Jr., Yao, J. and Wright, P. E. (1998) Sequence requirements for stabilization of a peptide reverse turn in water solution. Proline is not essential for stability. Eur. J. Biochem. 255, 462-471

Desai, U. R., Osterhout, J. J. and Klibanov, A. M. (1994). Protein Structure in the Lyophilized State: A Hydrogen Isotope Exchange/NMR Study with Bovine Pancreatic Trypsin Inhibitor. J. Am. Chem. Soc. 116, 9420-9422

Osterhout, J. J. Jr., Handel, T., Na, G.,Toumadje, A., Long, R. C., Connolly, P. J., Hoch, J. C., Johnson, W. C. Jr., Live, D. and DeGrado, W. F. (1992). Characterization of the Structural Properties of a1B, a Peptide Designed to Form a Four-Helix Bundle. J. Am. Chem. Soc. 114, 331-337

Osterhout, J. J. Jr., Baldwin, R. L., York, E. J., Stewart, J. M., Dyson, H. J. and Wright, P. E. (1989). 1H NMR Studies of the Solution Conformations of an Analogue of the C-Peptide of Ribonuclease A. Biochemistry 28, 7059-7064

Nall, B. T., Osterhout, J. J., Jr. and Ramdas, L. (1988). pH Dependence of Folding of Iso-2-cytochrome c. Biochemistry 27, 7310-7314

Osterhout, J. J. Jr., Muthukrishnan, K. and Nall, B. T. (1985). pH-Induced Conformation Changes and Equilibrium Unfolding in Yeast Iso-2 Cytochrome c. Biochemistry 24, 6680-6684

Osterhout, J. J., Jr. and Nall, B. T. (1985). Slow Refolding Kinetics in Yeast Iso-2-Cytochrome c. Biochemistry 24, 7999-8005

Other Publications

Osterhout, J. J., Jr., Lax, S. R. and Ravel, J. M. (1983). Factors From Wheat Germ That Enhance the Activity of Eukaryotic Initiation Factor eIF-2: Isolation and Characterization of Co-eIF-2alpha. J. Biol. Chem.. 258, 8285-8289

Lax, S. R., Osterhout, J. J., Jr. and Ravel, J. M. (1982). Factors From Wheat Germ That Enhance the Activity of Eukaryotic Initiation Factor eIF-2: Isolation and Characterization of Co-eIF-2beta. J. Biol. Chem.. 247, 8233-8237

Osterhout, J. J. Jr. and Alexanian, R. (1974). Ferrokinetic Studies of Mouse Erythryopoiesis. Exprimental Hematology 2, 307

Dissertation:
Isolation and Characterization of Co-eIF-2alpha and Co-eIF-2beta, Two Proteins From Wheat Germ That Stimulate the Activity of Initiation Factor eIF-2.

Professional Experience/Education

Member, Rowland Institute for Science, 1992 to present
Postdoctoral Associate, Dr. Jeffry C. Hoch, Rowland Institute for Science, 1988 to 1992
Postdoctoral Associate, Dr. Peter E. Wright, Research Institute of Scripps Clinic, 1988
Postdoctoral Associate, Dr. Robert L. Baldwin, Stanford University, 1985 to 1988
Welch Fellow, Dr. Barry T. Nall, University of Texas Medical School at Houston, 1982 to 1985
Ph. D. Chemistry, Dr. Joanne M. Ravel, University of Texas at Austin, 1981
B. A. Biochemistry, Rice University, Houston, Texas, 1974

Folding and Design Laboratory Members John Osterhout Sunita Kulkarni Research alpha-t-alpha Peptide Dissection HX Group Alumni Youcef Fezoui Marcela Oslin Tanya Knubovets Diane Schaak Wujing Xian	John J. Osterhout Protein Folding and Design Laboratory Contact Information Research Associate Professor Institute for Biomedical Science and Biotechnology BSW 362 P.O. Box 210088 University of Arizona Tucson, Arizona 85721 Telephone: 520-626-1311 E-mail: johno@email.arizona.edu
	Research Interests Recent fast kinetic experiments show that proteins can fold rapidly (on the millisecond time scale) in the absence of kinetic blocks such as proline isomerization or disulfide formation. Both the old view of protein folding (intermediates/pathways) and the new view (ensembles/energy landscapes) provide suggestions about how proteins might dodge the Levinthal paradox and manage to attain their native conformations on time scales less than eons. However, no matter which view of protein folding one prefers, a correlation between structure, stability, and the manner in which short and long range interactions conspire to direct folding along a pathway or funnel toward the native state is still desirable. This is a difficult problem to attack experimentally because the interesting intermediates are only transiently populated in kinetic experiments and fractionally populated in equilibrium experiments. Our approach to this problem is to use peptides as model systems. Our primary interest is the study of folding at the level of secondary structure association. To this end, a 38-residue peptide, alpha-t-alpha was designed de novo to form a helical hairpin in solution. The success of this design was confirmed by NMR structure determination. Current research is directed toward understanding the balance of forces contributing to the stability of the molecule and toward measuring the kinetics of helix association.
	Publications Alpha-t-alpha Publications Fezoui, Y, Hartley, D. M., Walsh, D. M., Selkoe, D. J., Osterhout, J. J. and Teplow, D. B. (2000) A de novo designed helix-turn-helix peptide forms nontoxic amyloid fibrils. Nature Structural Biology 7, 1095-1099 Fezoui, Y., Braswell, E. H., Xian, W. and Osterhout, J. J. (1999) Dissection of the de Novo Designed Peptide alpha-t-alpha: Stability and Properties of the Intact Molecule and Its Constituent Helices. Biochemistry 38, 2796-2804 Fezoui, Y., Connolly, P. J. and Osterhout, J. J. (1997) Solution Structure of alpha-t-alpha, a Helical Hairpin Peptide of De Novo Design. Protein Science 6, 1869-1877 Fezoui, Y., Weaver, D. L. and Osterhout, J. J. (1995). Strategies and Rationales for the De Novo Design of a Helical Hairpin Peptide. Protein Science 4, 286-295 Fezoui, Y., Weaver, D. L. and Osterhout, J. J. (1994). De Novo Design and Structural Characterization of an alpha-Helical Hairpin Peptide - A Model System for the Study of Protein Folding Intermediates. Proc. Natl. Acad. Sci. USA 91, 3675-3679 Protein Folding Publications Knubovets, T., Osterhout, J. J., Connolly, P. J. and Klibanov, A. M. (1999). Structure, thermostability, and conformational flexibility of hen egg-white lysozyme dissolved in glycerol. Proc. Natl. Acad. Sci. USA 96, 1262-1267 Knubovets, T., Osterhout, J. J. and Klibanov, A. M. (1999). Structure of Lysozyme Dissolved in Neat Organic Solvents as Assessed by NMR and CD Spectroscopies. Biotechnology and Bioengineering 63, 242-248 Choe, S. E., Matsudaira, P. T., Osterhout, J., Wagner, G., and Shakhnovich, E. I., (1998). Folding Kinetics of Villin 14T, a Protein Domain with a Central beta-Sheet and Two Hydrophobic Cores. Biochemistry 37, 14508-14518 Dyson, H. J., Bolinger, L., Feher, V. A. Osterhout, J. J. Jr., Yao, J. and Wright, P. E. (1998) Sequence requirements for stabilization of a peptide reverse turn in water solution. Proline is not essential for stability. Eur. J. Biochem. 255, 462-471 Desai, U. R., Osterhout, J. J. and Klibanov, A. M. (1994). Protein Structure in the Lyophilized State: A Hydrogen Isotope Exchange/NMR Study with Bovine Pancreatic Trypsin Inhibitor. J. Am. Chem. Soc. 116, 9420-9422 Osterhout, J. J. Jr., Handel, T., Na, G.,Toumadje, A., Long, R. C., Connolly, P. J., Hoch, J. C., Johnson, W. C. Jr., Live, D. and DeGrado, W. F. (1992). Characterization of the Structural Properties of a1B, a Peptide Designed to Form a Four-Helix Bundle. J. Am. Chem. Soc. 114, 331-337 Osterhout, J. J. Jr., Baldwin, R. L., York, E. J., Stewart, J. M., Dyson, H. J. and Wright, P. E. (1989). 1H NMR Studies of the Solution Conformations of an Analogue of the C-Peptide of Ribonuclease A. Biochemistry 28, 7059-7064 Nall, B. T., Osterhout, J. J., Jr. and Ramdas, L. (1988). pH Dependence of Folding of Iso-2-cytochrome c. Biochemistry 27, 7310-7314 Osterhout, J. J. Jr., Muthukrishnan, K. and Nall, B. T. (1985). pH-Induced Conformation Changes and Equilibrium Unfolding in Yeast Iso-2 Cytochrome c. Biochemistry 24, 6680-6684 Osterhout, J. J., Jr. and Nall, B. T. (1985). Slow Refolding Kinetics in Yeast Iso-2-Cytochrome c. Biochemistry 24, 7999-8005 Other Publications Osterhout, J. J., Jr., Lax, S. R. and Ravel, J. M. (1983). Factors From Wheat Germ That Enhance the Activity of Eukaryotic Initiation Factor eIF-2: Isolation and Characterization of Co-eIF-2alpha. J. Biol. Chem.. 258, 8285-8289 Lax, S. R., Osterhout, J. J., Jr. and Ravel, J. M. (1982). Factors From Wheat Germ That Enhance the Activity of Eukaryotic Initiation Factor eIF-2: Isolation and Characterization of Co-eIF-2beta. J. Biol. Chem.. 247, 8233-8237 Osterhout, J. J. Jr. and Alexanian, R. (1974). Ferrokinetic Studies of Mouse Erythryopoiesis. Exprimental Hematology 2, 307 Dissertation: Isolation and Characterization of Co-eIF-2alpha and Co-eIF-2beta, Two Proteins From Wheat Germ That Stimulate the Activity of Initiation Factor eIF-2.
	Professional Experience/Education Member, Rowland Institute for Science, 1992 to present Postdoctoral Associate, Dr. Jeffry C. Hoch, Rowland Institute for Science, 1988 to 1992 Postdoctoral Associate, Dr. Peter E. Wright, Research Institute of Scripps Clinic, 1988 Postdoctoral Associate, Dr. Robert L. Baldwin, Stanford University, 1985 to 1988 Welch Fellow, Dr. Barry T. Nall, University of Texas Medical School at Houston, 1982 to 1985 Ph. D. Chemistry, Dr. Joanne M. Ravel, University of Texas at Austin, 1981 B. A. Biochemistry, Rice University, Houston, Texas, 1974