Designing heterodimeric two-stranded alpha-helical coiled-coils: the effect of chain length on protein folding, stability and specificity.
The E/K coil, a heterodimeric coiled-coil, has been designed as a universal peptide capture and delivery system for use in applications such as biosensors and affinity chromatography. In this design, heterodimer formation is specified through the placement of charged residues at the e and g positions of the heptad repeat. The affinity and stability of the E/K coil has been modified in order to allow a greater range of conditions for association and dissociation by varying the chain length to obtain three, four and five heptad coiled-coils (21, 28 and 35 residues per polypeptide chain). The effect of chain length on stability and folding was examined by circular dichroism spectroscopy, guanidine hydrochloride denaturation, and redox equilibrium experiments. It was found that increases in chain length produced increases in the stability of heterodimeric coiled-coils, but in a nonlinear fashion. The resulting disulfide-bridged heterostranded molecules and reduced heterodimers span a wide range of stabilities (deltaG=3.3-11.9 kcal/mol), greatly expanding their scope for use in protein engineering and biomedical applications.
Litowski
J R
JR
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Hodges
R S
RS
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D019281:Dimerization; D019791:Guanidine / Q000737:chemistry; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*
D011506:Proteins; D019791:Guanidine