Abstract

This is a renewal proposal (years 5-10) to continue studies of the folding and assembly mechanisms in multisubunit proteins. Specifically, the questions to be addressed are: (1) What are the important structural features of the transient intermediates and intervening transition states observed during the folding of specific representative multimeric protein? (2) Does pre-formed structure in the monomer affect the folding kinetics of the dimer? (3) Do partially folded associated species guide the refolding reaction to the native dimer (trimer)? A combination of biophysical techniques spanning mutagenesis, hydrogen-deuterium exchange, NMR, Mass Spectrometry and rapid-mixing optical techniques will be used to gain information on the time-dependent structure formation and association reactions during the folding of three distinct systems: (a) coiled-coils, (b) the trp-repressor and (c) the trimeric beta-helix proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM054836-05S1
Application #
6333307
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Program Officer
Wehrle, Janna P
Project Start
1996-07-01
Project End
2001-06-30
Budget Start
2000-09-01
Budget End
2001-06-30
Support Year
5
Fiscal Year
2000
Total Cost
$41,182
Indirect Cost

  Institution

Name
Pennsylvania State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
University Park
State
PA
Country
United States
Zip Code
16802

  Publications

Niu, Ben; Mackness, Brian C; Rempel, Don L et al. (2017) Incorporation of a Reporter Peptide in FPOP Compensates for Adventitious Scavengers and Permits Time-Dependent Measurements. J Am Soc Mass Spectrom 28:389-392
Kathuria, Sagar V; Chan, Yvonne H; Nobrega, R Paul et al. (2016) Clusters of isoleucine, leucine, and valine side chains define cores of stability in high-energy states of globular proteins: Sequence determinants of structure and stability. Protein Sci 25:662-75
Boopathy, Sivakumar; Silvas, Tania V; Tischbein, Maeve et al. (2015) Structural basis for mutation-induced destabilization of profilin 1 in ALS. Proc Natl Acad Sci U S A 112:7984-9
Mackness, Brian C; Tran, Meme T; McClain, Shannan P et al. (2014) Folding of the RNA recognition motif (RRM) domains of the amyotrophic lateral sclerosis (ALS)-linked protein TDP-43 reveals an intermediate state. J Biol Chem 289:8264-76
Kathuria, Sagar V; Kayatekin, Can; Barrea, Raul et al. (2014) Microsecond barrier-limited chain collapse observed by time-resolved FRET and SAXS. J Mol Biol 426:1980-94
Kayatekin, Can; Cohen, Noah R; Matthews, C Robert (2012) Enthalpic barriers dominate the folding and unfolding of the human Cu, Zn superoxide dismutase monomer. J Mol Biol 424:192-202
Kathuria, Sagar V; Guo, Liang; Graceffa, Rita et al. (2011) Minireview: structural insights into early folding events using continuous-flow time-resolved small-angle X-ray scattering. Biopolymers 95:550-8
Svensson, Anna-Karin E; Bilsel, Osman; Kayatekin, Can et al. (2010) Metal-free ALS variants of dimeric human Cu,Zn-superoxide dismutase have enhanced populations of monomeric species. PLoS One 5:e10064
Kayatekin, Can; Zitzewitz, Jill A; Matthews, C Robert (2010) Disulfide-reduced ALS variants of Cu, Zn superoxide dismutase exhibit increased populations of unfolded species. J Mol Biol 398:320-31
Tiwari, Ashutosh; Liba, Amir; Sohn, Se Hui et al. (2009) Metal deficiency increases aberrant hydrophobicity of mutant superoxide dismutases that cause amyotrophic lateral sclerosis. J Biol Chem 284:27746-58

Showing the most recent 10 out of 27 publications