Solubilizing Mutations Used to Crystallize One CFTR Domain Attenuate the Trafficking and Channel Defects Caused by the Major Cystic Fibrosis Mutation

Luísa S. Pissarra, Carlos M. Farinha, Zhe Xu, André Schmidt, Patrick H. Thibodeau, Zhiwei Cai, Philip J Thomas, David N. Sheppard, Margarida D. Amaral

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) Cl- channel. F508del, the most frequent CF-causing mutation, disrupts both the processing and function of CFTR. Recently, the crystal structure of the first nucleotide-binding domain of CFTR bearing F508del (F508del-NBD1) was elucidated. Although F508del-NBD1 shows only minor conformational changes relative to that of wild-type NBD1, additional mutations (F494N/Q637R or F429S/F494N/Q637R) were required for domain solubility and crystallization. Here we show that these solubilizing mutations in cis with F508del partially rescue the trafficking defect of full-length F508del-CFTR and attenuate its gating defect. We interpret these data to suggest that the solubilizing mutations utilized to facilitate F508del-NBD1 production also assist folding of full-length F508del-CFTR protein. Thus, the available crystal structure of F508del-NBD1 might correspond to a partially corrected conformation of this domain.

Original languageEnglish (US)
Pages (from-to)62-69
Number of pages8
JournalChemistry and Biology
Volume15
Issue number1
DOIs
StatePublished - Jan 25 2008

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Cystic Fibrosis
Bearings (structural)
Crystal structure
Defects
Mutation
Crystallization
Conformations
Nucleotides
Solubility
Processing
Proteins

Keywords

  • CELLBIO
  • CHEMBIO

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Solubilizing Mutations Used to Crystallize One CFTR Domain Attenuate the Trafficking and Channel Defects Caused by the Major Cystic Fibrosis Mutation. / Pissarra, Luísa S.; Farinha, Carlos M.; Xu, Zhe; Schmidt, André; Thibodeau, Patrick H.; Cai, Zhiwei; Thomas, Philip J; Sheppard, David N.; Amaral, Margarida D.

In: Chemistry and Biology, Vol. 15, No. 1, 25.01.2008, p. 62-69.

Research output: Contribution to journalArticle

Pissarra, Luísa S. ; Farinha, Carlos M. ; Xu, Zhe ; Schmidt, André ; Thibodeau, Patrick H. ; Cai, Zhiwei ; Thomas, Philip J ; Sheppard, David N. ; Amaral, Margarida D. / Solubilizing Mutations Used to Crystallize One CFTR Domain Attenuate the Trafficking and Channel Defects Caused by the Major Cystic Fibrosis Mutation. In: Chemistry and Biology. 2008 ; Vol. 15, No. 1. pp. 62-69.
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AB - Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) Cl- channel. F508del, the most frequent CF-causing mutation, disrupts both the processing and function of CFTR. Recently, the crystal structure of the first nucleotide-binding domain of CFTR bearing F508del (F508del-NBD1) was elucidated. Although F508del-NBD1 shows only minor conformational changes relative to that of wild-type NBD1, additional mutations (F494N/Q637R or F429S/F494N/Q637R) were required for domain solubility and crystallization. Here we show that these solubilizing mutations in cis with F508del partially rescue the trafficking defect of full-length F508del-CFTR and attenuate its gating defect. We interpret these data to suggest that the solubilizing mutations utilized to facilitate F508del-NBD1 production also assist folding of full-length F508del-CFTR protein. Thus, the available crystal structure of F508del-NBD1 might correspond to a partially corrected conformation of this domain.

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