title: Effect of chain length on coiled-coil stability: decreasing stability with increasing chain length.
abstract: The de novo design and biophysical characterization of three series of two-stranded alpha-helical coiled coils with different chain lengths are described. Our goal was to examine how increasing chain length would affect protein folding and stability when one or more heptad repeat(s) of K-A-E-A-L-E-G (gabcdef) was inserted into the central region of different coiled-coil host proteins. This heptad was designed to maintain the continuous 3-4 hydrophobic repeat of the coiled-coil host and introduce an Ala and Leu residue in the hydrophobic core at the a and d position, respectively, and a pair of stabilizing interchain ionic i to i' + 5 (g to e') interactions per heptad inserted. The secondary structures of the three series of disulfide-bridged polypeptides were studied by CD spectroscopy and their stabilities determined by chemical and thermal denaturation. The results showed that successive insertions of this heptad systematically decreased the stability of all the coiled coils studied regardless of the overall initial stability of the host coiled coil. These observations are in contrast to the generally accepted implication that the folding and stability of coiled coils are enhanced with increasing chain length. Our results imply that, in these examples where an Ala and Leu hydrophobic residue were introduced into the coiled-coil core per inserted heptad, there was still insufficient stability to overcome unfavorable entropy associated with chain length extension, even though the inserted heptad contained the most stabilizing hydrophobic residue (Leu) at position d and stabilizing ionic attractions.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center at Fitzsimons, Aurora, CO 80045-0511, USA.
authors.forename: Stanley C
authors.initials: SC
authors.lastname: Kwok
authors.forename: Robert S
authors.initials: RS
authors.lastname: Hodges
mesh_terms: D010455:Peptides / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D013696:Temperature; D014508:Urea
chemical_list: D010455:Peptides; D011506:Proteins; D014508:Urea