Premium
α‐helix to random coil transitions: Determination of peptide concentration from the CD at the isodichroic point
Author(s) -
Holtzer Marilyn Emerson,
Holtzer Alfred
Publication year - 1992
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.360321209
Subject(s) - chemistry , lambda , random coil , peptide , helix (gastropod) , crystallography , population , transition point , analytical chemistry (journal) , physics , chromatography , circular dichroism , thermodynamics , optics , ecology , biochemistry , demography , sociology , snail , biology
A method is presented for determining the concentrations of peptides and proteins having isodichroic points near 203 nm. The existence of an isodichroic point for a given substance indicates a local two‐state (α‐helix, random coil) population. The mean residue ellipticity at the isodichroic point, [θ λ i ], is, of course, independent of helix content. For a wide variety of synthetic and natural peptides, including both single helices and coiled coils, it is shown that [θ λ i ] is also essentially independent of substance and of whether the transition is induced by temperature, ionic strength, pH, chain length changes, amino acid substitution, or solvent perturbation. Averaging [θ λ i ] values culled from various laboratories gives –151 ± 16 (SD, 7 sources) deg · cm 2 · mmol −1 . In our laboratory, nonpolymerizable rabbit α‐tropomyosin and two α‐tropomyosin subsequences yield –135 ± 10 (SD, 190 values) deg · cm 2 · mmol −1 . Thus, given [θ λ i ] for a peptide of known concentration, it is possible to estimate the concentration of any other peptide provided that it has an isodichroic point at which the ellipticity is accurately measurable. It is then possible to calculate [θ λ ] at any other wavelength for which θ is known. It is advisable to determine [θ λ i ] for the best known peptide in one's own laboratory, since it depends on absolute instrument and cell calibrations and an absolute concentration determination. © 1992 John Wiley & Sons, Inc.