Cystic fibrosis transmembrane conductance regulator: Solution structures of peptides based on the Phe508 region, the most common site of disease- causing ΔF508 mutation

Michael A. Massiah, Young Hee Ko, Peter L. Pedersen, Albert S. Mildvan

Research output: Contribution to journalArticlepeer-review

Abstract

Most cases of cystic fibrosis (CF), a common inherited disease of epithelial cell origin, are caused by the deletion of Phe508 located in the first nucleotide-binding domain (NBF1) of the protein called CFTR (cystic fibrosis transmembrane conductance regulator). To gain greater insight into the structure within the Phe508 region of the wild-type protein and the change in structure that occurs when this residue is deleted, we conducted nuclear magnetic resonance (NMR) studies on representative synthetic 26 and 25 amino acid peptide segments. 2D 1H NMR studies at 600 MHz of the 26- residue peptide consisting of Met498 to Ala523 in 10% DMSO, pH 4.0, at 25 °C show a continuous but labile helix from Gly500 to Lys522, based on both NH- NH((i.i+1)) and αH-NH((i,i+1)) NOEs. Phe508 within this helix shows only short-range (i, ≤i + 2) NOEs. The corresponding 25-residue peptide lacking Phe508 also forms a labile helix from Gly500 to Lys522. However, the relative intensities of the NH-NH((i,i+1))/αHNH((i,i+i)) NOEs, fewer intermediate- range NOEs, and downfield αH and NH chemical shifts indicate a lower helical propensity of the 25-mer between residues 505 and 517, surrounding the missing residue, Phe508.2D 1H NMR studies of both peptides in saturating (43%) TFE reveal stable α-helices from Gly500 to Lys522, based on NH- NH((i,i+1,2,3)), αHNH((i.i+2,3,4)), αH-β((i,i+3)), and weak αH- NH((i,i+1)) NOEs. However, downfield shifts of the αH resonances from residues Gly500 to Ile507 and fewer intermediate-range NOEs suggest a less stable α-helix in the 25-mer even in saturating TFE. These findings show that the Phe508-containing region of CFTR has a propensity to form an α- helix, which is destabilized by the ΔF508 mutation found in most patients with CF. These studies have direct relevance to better understanding the CFTR misfolding problem associated with CF and to identifying chemical agents, which correct this problem.

Original languageEnglish (US)
Pages (from-to)7453-7461
Number of pages9
JournalBiochemistry
Volume38
Issue number23
DOIs
StatePublished - Jun 8 1999

ASJC Scopus subject areas

  • Biochemistry

Fingerprint

Dive into the research topics of 'Cystic fibrosis transmembrane conductance regulator: Solution structures of peptides based on the Phe508 region, the most common site of disease- causing ΔF508 mutation'. Together they form a unique fingerprint.

Cite this