Sy. Venyaminov et al., Analysis of the segmental stability of helical peptides by isotope-edited infrared spectroscopy, PROTEINS, 45(1), 2001, pp. 81-89
Isotope-edited infrared spectroscopy has the ability to probe the segmental
properties of long biopolymers. In this work, we have compared the infrare
d spectra of a model helical peptide (C-12) Ac-W-(E-A-A-A-R)(6)-A-NH2, desc
ribed originally by Merutka et al. (Biochemistry 1991;30: 4245-4248) and th
ree derivatives that are C-13 labeled at the backbone carbonyl of alanines.
The locations of six isotopically labeled alanines are at the N-terminal,
C-terminal, and the middle two repeating units of the peptide. Variation in
temperature from 1 degrees to 91 degreesC transformed the peptides from pr
edominantly helical to predominantly disordered state. Amplitude and positi
on of the infrared amide I' absorption bands from C-12- and C-13-labeled se
gments provided information about the helical content. Temperature dependen
ce of infrared spectra was used to estimate segmental stability. As a contr
ol measure of overall peptide stability and helicity (independent of labeli
ng), the temperature dependence of circular dichroism spectra in the far-UV
range at identical conditions (temperature and solvent) as infrared spectr
a was measured. The results indicate that the central quarter of the 32 ami
no acids helix has the maximal helicity and stability. The midpoint of the
melting curve of the central quarter of the helix is 5.4 +/- 0.8 degreesC h
igher than that of the termini. The N-terminal third of the helix is more h
elical and is 2.0 +/- 1.4 degreesC more stable than the C-terminus. (C) 200
1 Wiley-Liss, Inc.