A small molecule that binds to an ATPase domain of Hsc70 promotes membrane trafficking of mutant cystic fibrosis transmembrane conductance regulator

Hyungseoph J. Cho, Heon Yung Gee, Kyung Hwa Baek, Sung Kyun Ko, Jong Moon Park, Hookeun Lee, Nam Doo Kim, Min Goo Lee, Injae Shin

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Cystic fibrosis transmembrane conductance regulator (CFTR) is a cell-surface anion channel that permeates chloride and bicarbonate ions. The most frequent mutation of CFTR that causes cystic fibrosis is the deletion of phenylalanine at position 508 (ΔF508), which leads to defects in protein folding and cellular trafficking to the plasma membrane. The lack of the cell-surface CFTR results in a reduction in the lifespan due to chronic lung infection with progressive deterioration of lung function. Hsc70 plays a crucial role in degradation of mutant CFTR by the ubiquitin-proteasome system. To date, various Hsc70 inhibitors and transcription regulators have been tested to determine whether they correct the defective activity of mutant CFTR. However, they exhibited limited or questionable effects on restoring the chloride channel activity in cystic fibrosis cells. Herein, we show that a small molecule apoptozole (Az) has high cellular potency to promote membrane trafficking of mutant CFTR and its chloride channel activity in cystic fibrosis cells. Results from affinity chromatography and ATPase activity assay indicate that Az inhibits the ATPase activity of Hsc70 by binding to its ATPase domain. In addition, a ligand-directed protein labeling and molecular modeling studies also suggest the binding of Az to an ATPase domain, in particular, an ATP-binding pocket. It is proposed that Az suppresses ubiquitination of ΔF508-CFTR maybe by blocking interaction of the mutant with Hsc70 and CHIP, and, as a consequence, it enhances membrane trafficking of the mutant.

Original languageEnglish
Pages (from-to)20267-20276
Number of pages10
JournalJournal of the American Chemical Society
Volume133
Issue number50
DOIs
Publication statusPublished - 2011 Dec 21

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Cystic Fibrosis Transmembrane Conductance Regulator
Adenosine Triphosphatases
Membranes
Molecules
Chloride Channels
Cystic Fibrosis
Affinity chromatography
Protein folding
Molecular modeling
Adenosinetriphosphate
Transcription
Cell membranes
Labeling
Deterioration
Assays
Negative ions
Ligands
Lung
Proteins
Ubiquitination

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Cho, Hyungseoph J. ; Gee, Heon Yung ; Baek, Kyung Hwa ; Ko, Sung Kyun ; Park, Jong Moon ; Lee, Hookeun ; Kim, Nam Doo ; Lee, Min Goo ; Shin, Injae. / A small molecule that binds to an ATPase domain of Hsc70 promotes membrane trafficking of mutant cystic fibrosis transmembrane conductance regulator. In: Journal of the American Chemical Society. 2011 ; Vol. 133, No. 50. pp. 20267-20276.
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abstract = "Cystic fibrosis transmembrane conductance regulator (CFTR) is a cell-surface anion channel that permeates chloride and bicarbonate ions. The most frequent mutation of CFTR that causes cystic fibrosis is the deletion of phenylalanine at position 508 (ΔF508), which leads to defects in protein folding and cellular trafficking to the plasma membrane. The lack of the cell-surface CFTR results in a reduction in the lifespan due to chronic lung infection with progressive deterioration of lung function. Hsc70 plays a crucial role in degradation of mutant CFTR by the ubiquitin-proteasome system. To date, various Hsc70 inhibitors and transcription regulators have been tested to determine whether they correct the defective activity of mutant CFTR. However, they exhibited limited or questionable effects on restoring the chloride channel activity in cystic fibrosis cells. Herein, we show that a small molecule apoptozole (Az) has high cellular potency to promote membrane trafficking of mutant CFTR and its chloride channel activity in cystic fibrosis cells. Results from affinity chromatography and ATPase activity assay indicate that Az inhibits the ATPase activity of Hsc70 by binding to its ATPase domain. In addition, a ligand-directed protein labeling and molecular modeling studies also suggest the binding of Az to an ATPase domain, in particular, an ATP-binding pocket. It is proposed that Az suppresses ubiquitination of ΔF508-CFTR maybe by blocking interaction of the mutant with Hsc70 and CHIP, and, as a consequence, it enhances membrane trafficking of the mutant.",
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A small molecule that binds to an ATPase domain of Hsc70 promotes membrane trafficking of mutant cystic fibrosis transmembrane conductance regulator. / Cho, Hyungseoph J.; Gee, Heon Yung; Baek, Kyung Hwa; Ko, Sung Kyun; Park, Jong Moon; Lee, Hookeun; Kim, Nam Doo; Lee, Min Goo; Shin, Injae.

In: Journal of the American Chemical Society, Vol. 133, No. 50, 21.12.2011, p. 20267-20276.

Research output: Contribution to journalArticle

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T1 - A small molecule that binds to an ATPase domain of Hsc70 promotes membrane trafficking of mutant cystic fibrosis transmembrane conductance regulator

AU - Cho, Hyungseoph J.

AU - Gee, Heon Yung

AU - Baek, Kyung Hwa

AU - Ko, Sung Kyun

AU - Park, Jong Moon

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AU - Kim, Nam Doo

AU - Lee, Min Goo

AU - Shin, Injae

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