eprintid: 17791
rev_number: 22
userid: 1419
dir: disk0/00/01/77/91
datestamp: 2013-03-20 16:43:38
lastmod: 2019-02-02 16:55:12
status_changed: 2013-03-20 16:43:38
type: article
metadata_visibility: show
item_issues_count: 0
eprint_status: archive
creators_name: Lednev, IK
creators_name: Karnoup, AS
creators_name: Sparrow, MC
creators_name: Asher, SA
creators_email: 
creators_email: 
creators_email: 
creators_email: asher@pitt.edu
creators_id: 
creators_id: 
creators_id: 
creators_id: ASHER
title: α-Helix peptide folding and unfolding activation barriers: A nanosecond UV resonance raman study
ispublished: pub
divisions: sch_as_chemistry
full_text_status: public
abstract: We used UV resonance Raman spectroscopy to characterize the equilibrium conformation and the kinetics of thermal denaturation of a 21 amino acid, mainly alanine, α-helical peptide (AP). The 204-nm UV resonance Raman spectra show selective enhancements of the amide vibrations, whose intensities and frequencies strongly depend on the peptide secondary structure. These AP Raman spectra were accurately modeled by a linear combination of the temperature-dependent Raman spectra of the pure random coil and the pure α-helix conformations; this demonstrates that the AP helix-coil equilibrium is well-described by a two-state model. We constructed a new transient UV resonance Raman spectrometer and developed the necessary methodologies to measure the nanosecond relaxation of AP following a 3-ns T- jump. We obtained the T-jump by using a 1.9-μm IR pulse that heats the solvent water. We probed the AP relaxation using delayed 204-nm excitation pulses which excite the Raman spectra of the amide backbone vibrations. We observe little AP structural changes within the first 40 ns, after which the α-helix starts unfolding. We determined the temperature dependence of the folding and unfolding rates and found that the unfolding rate constants show Arrheniustype behavior with an apparent κ8 kcal/mol activation barrier and a reciprocal rate constant of 240 ± 60 ns at 37 °C. However, the folding rate constants show a negative activation barrier, indicating a failure of transitionstate theory in the simple two-state modeling of AP thermal unfolding, which assumes a temperature-independent potential energy profile along the reaction coordinate. Our measurements of the initial steps in the α-helical structure evolution support recent protein folding landscape and funnel theories; our temperature-dependent rate constants sense the energy landscape complexity at the earliest stages of folding and unfolding.
date: 1999-09-08
date_type: published
publication: Journal of the American Chemical Society
volume: 121
number: 35
pagerange: 8074 - 8086
refereed: TRUE
issn: 0002-7863
id_number: 10.1021/ja991382f
citation:   Lednev, IK and Karnoup, AS and Sparrow, MC and Asher, SA  (1999) α-Helix peptide folding and unfolding activation barriers: A nanosecond UV resonance raman study.  Journal of the American Chemical Society, 121 (35).  8074 - 8086.  ISSN 0002-7863     
document_url: http://d-scholarship-dev.library.pitt.edu/17791/1/licence.txt