Evidence for a partially structured state of the amylin monomer.
Islet amyloid polypeptide (amylin) is the main component in amyloid deposits formed in type II diabetes. We used triplet quenching to probe the dynamics of contact formation between the N-terminal disulfide loop and a C-terminal tryptophan in monomeric amylins from human and rat. Quenching rates measured in the absence of denaturant are four times larger than those in 6 M guanidinium chloride, indicating a decrease in the average end-to-end distance (collapse) at low denaturant concentrations. We were surprised to find an even greater (sevenfold) increase in quenching rates on removal of denaturant for a hydrophilic control peptide containing the disulfide loop compared to the same peptide without the loop (twofold change). These results suggest that collapse is driven by backbone-backbone and backbone-side chain interactions involving the disulfide loop portion of the chain rather than by the formation of side-chain hydrophobic contacts. Molecular dynamics simulations of the control peptide show that the collapse results from hydrogen-bonding interactions between the central residues of the chain and the disulfide loop. The quenching experiments also indicate that the monomer of the human, amyloidogenic form of amylin is more compact than the rat form, which does not form amyloid. We discuss these newly observed differences between human and rat amylin in solution and their possible relation to aggregation and to the physiological function of amylin binding to the calcitonin receptor.
Vaiana
Sara M
SM
Laboratory of Chemical Physics, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA. sara.vaiana@asu.edu
Best
Robert B
RB
Yau
Wai-Ming
WM
Eaton
William A
WA
Hofrichter
James
J
D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D004058:Diffusion; D004220:Disulfides / Q000737:chemistry; D019791:Guanidine / Q000494:pharmacology; D006801:Humans; D058228:Islet Amyloid Polypeptide; D007700:Kinetics; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D051381:Rats; D012015:Reference Standards; D014364:Tryptophan; D014867:Water / Q000737:chemistry
D000682:Amyloid; D004220:Disulfides; D058228:Islet Amyloid Polypeptide; D010455:Peptides; D014867:Water; D014364:Tryptophan; D019791:Guanidine