Ligand-Driven Vectorial Folding of Ribosome-Bound Human CFTR NBD1

Amardeep Khushoo, Zhongying Yang, Arthur E. Johnson, William Skach

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The mechanism by which protein folding is coupled to biosynthesis is a critical, but poorly understood, aspect of protein conformational diseases. Here we use fluorescence resonance energy transfer (FRET) to characterize tertiary structural transitions of nascent polypeptides and show that the first nucleotide-binding domain (NBD1) of human CFTR, whose folding is defective in cystic fibrosis, folds via a cotranslational multistep pathway as it is synthesized on the ribosome. Folding begins abruptly as NBD1 residues 389-500 emerge from the ribosome exit tunnel, initiating compaction of a small, N-terminal α/β-subdomain. Real-time kinetics of synchronized nascent chains revealed that subdomain folding is rapid, occurs coincident with synthesis, and is facilitated by direct ATP binding to the nascent polypeptide. These findings localize the major CF defect late in the NBD1 folding pathway and establish a paradigm wherein a cellular ligand promotes vectorial domain folding by facilitating an energetically favored local peptide conformation.

Original languageEnglish (US)
Pages (from-to)682-692
Number of pages11
JournalMolecular Cell
Volume41
Issue number6
DOIs
StatePublished - Mar 18 2011

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Ribosomes
Ligands
Peptides
Fluorescence Resonance Energy Transfer
Protein Folding
Cystic Fibrosis
Nucleotides
Adenosine Triphosphate
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Ligand-Driven Vectorial Folding of Ribosome-Bound Human CFTR NBD1. / Khushoo, Amardeep; Yang, Zhongying; Johnson, Arthur E.; Skach, William.

In: Molecular Cell, Vol. 41, No. 6, 18.03.2011, p. 682-692.

Research output: Contribution to journalArticle

Khushoo, A, Yang, Z, Johnson, AE & Skach, W 2011, 'Ligand-Driven Vectorial Folding of Ribosome-Bound Human CFTR NBD1', Molecular Cell, vol. 41, no. 6, pp. 682-692. https://doi.org/10.1016/j.molcel.2011.02.027
Khushoo, Amardeep ; Yang, Zhongying ; Johnson, Arthur E. ; Skach, William. / Ligand-Driven Vectorial Folding of Ribosome-Bound Human CFTR NBD1. In: Molecular Cell. 2011 ; Vol. 41, No. 6. pp. 682-692.
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