Abstract

This is a competitive renewal request for continuation of a program of studies aimed at understanding the chemical principals governing the folding, stability and self-association of beta-sheet structures in aqueous solution. Three lines of investigation are proposed: 1) the design and synthesis of templates that can be inserted into sequences to facilitate sheet formation and to determine of what kind of amino acids are thermodynamically favored to form sheets, and the role of hydrophobic clusters and hydrogen bonds in driving sheet formation; 2) studies of a naturally occurring 38-residue, 3-stranded antiparallel beta-sheet domain (termed the WW domain because of its 2 Trp residues) aimed at understanding the interactions that stabilize the domain structure; 3) studies of intermolecular beta-sheet formation using hosts that can selectively bind peptides. The peptides are obtained by solid phase chemical synthesis, and in the case of the WW domain, using a bacterial expression system. The peptide structures are evaluated by analytical ultracentrifugation, CD, FTIR, NMR, mass spectroscopy, binding of hydrophobic probes (ANS), fluorescence, as well as sequence analysis, in the case of the WW domain motif that has been identified in a number of proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM051105-07
Application #
6180374
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Wehrle, Janna P
Project Start
1994-09-01
Project End
2002-06-30
Budget Start
2000-09-01
Budget End
2002-06-30
Support Year
7
Fiscal Year
2000
Total Cost
$222,299
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Ardejani, Maziar S; Powers, Evan T; Kelly, Jeffery W (2017) Using Cooperatively Folded Peptides To Measure Interaction Energies and Conformational Propensities. Acc Chem Res 50:1875-1882
Chen, Wentao; Kong, Leopold; Connelly, Stephen et al. (2016) Stabilizing the CH2 Domain of an Antibody by Engineering in an Enhanced Aromatic Sequon. ACS Chem Biol 11:1852-61
Dave, Kapil; Jäger, Marcus; Nguyen, Houbi et al. (2016) High-Resolution Mapping of the Folding Transition State of a WW Domain. J Mol Biol 428:1617-36
Hsu, Che-Hsiung; Park, Sangho; Mortenson, David E et al. (2016) The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. J Am Chem Soc 138:7636-48
Chen, Wentao; Dong, Jiajia; Li, Suhua et al. (2016) Synthesis of Sulfotyrosine-Containing Peptides by Incorporating Fluorosulfated Tyrosine Using an Fmoc-Based Solid-Phase Strategy. Angew Chem Int Ed Engl 55:1835-8
Murray, Amber N; Chen, Wentao; Antonopoulos, Aristotelis et al. (2015) Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. Chem Biol 22:1052-62
Hebert, Daniel N; Lamriben, Lydia; Powers, Evan T et al. (2014) The intrinsic and extrinsic effects of N-linked glycans on glycoproteostasis. Nat Chem Biol 10:902-10
Chen, Wentao; Enck, Sebastian; Price, Joshua L et al. (2013) Structural and energetic basis of carbohydrate-aromatic packing interactions in proteins. J Am Chem Soc 135:9877-84
Price, Joshua L; Shental-Bechor, Dalit; Dhar, Apratim et al. (2012) Correction to Context-Dependent Effects of Asparagine Glycosylation on Pin WW Folding Kinetics and Thermodynamics. J Am Chem Soc 134:4450-4451
Araya, Carlos L; Fowler, Douglas M; Chen, Wentao et al. (2012) A fundamental protein property, thermodynamic stability, revealed solely from large-scale measurements of protein function. Proc Natl Acad Sci U S A 109:16858-63

Showing the most recent 10 out of 49 publications