A comparison of three- and four-helix bundle TASP molecules.
We have designed, synthesized and characterized three- and four-helix bundle template-assembled synthetic proteins CTASPs). The TASPs were synthesized using disulphide bonds between the peptides and either the cyclotribenzylene (CTB) template, or the cavitand (BOWL) template, to form the three- and four helix bundles, respectively. The TASPs were constructed using peptides that were linked via their N-termini (peptide CGGGEELLKKXEELLKKG, where X = L, I, Nle or V), or via their C-termini (peptide GEELLKKLEELLKKGGGC). Each TASP was assayed for its structure, stability, 'native-like' characteristics and whether it was a monomer in solution. All TASPs were found to be highly helical, and highly resistant to chemical denaturation using guanidine hydrochloride (GnHCl). Analysis of the GnHCl-induced unfolding curves of the different TASPs demonstrated stability differences based on the number of helices in the bundle, the end of the helix that was attached to the template, and the identity of the core amino acid. The TASPs all had molten-globule structure, which is (generally) consistent with a degenerate sequence in the core. The four-helix bundle TASPs appeared to be monomers in solution, whereas there is some evidence that the three-helix bundle TASPs are weakly self-associating.
Causton
Ashley S
AS
Department of Chemistry, Vancouver, BC, Canada.
Sherman
John C
JC
D000595:Amino Acid Sequence; D002499:Centrifugation, Density Gradient / Q000379:methods; D002942:Circular Dichroism; D004220:Disulfides / Q000737:chemistry; D004988:Ethers, Cyclic / Q000737:chemistry; D019791:Guanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D017711:Nonlinear Dynamics; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation / Q000187:drug effects; D017433:Protein Structure, Secondary; D011506:Proteins / Q000032:analysis / Q000138:chemical synthesis / Q000737:chemistry*; D012118:Resorcinols / Q000737:chemistry; D013816:Thermodynamics
D004220:Disulfides; D004988:Ethers, Cyclic; D011506:Proteins; D012118:Resorcinols; C442939:cavitand; D019791:Guanidine