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Alteration of CFTR transmembrane span integration by disease-causing mutations

Anna E Patrick1, Andrey L Karamyshev, Linda Millen

  • 1Department of Physiology, University of Texas Southwestern Medical Center, Dallas, TX 75235, USA.

Molecular Biology of the Cell
|October 15, 2011
PubMed

Insights

Cystic fibrosis mutations in CFTR protein spans cause misfolding and ER accumulation. Different mutations, like G85E and G91R, lead to distinct molecular disruptions, explaining CF pathology.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Missense mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) protein are a primary cause of cystic fibrosis (CF).
  • Many CF-causing CFTR mutations lead to protein misfolding and accumulation in the endoplasmic reticulum (ER), preventing proper protein maturation and function.
  • Specific mutations within CFTR's transmembrane (TM) domains are implicated in altered protein integration, but their precise effects remain unclear.

Purpose of the Study:

  • To experimentally determine the ER luminal integration profiles of CFTR's first two transmembrane spans (TM1 and TM2).
  • To investigate the impact of two CF-associated missense mutations, G85E and G91R, located in TM1, on CFTR's integration and conformation within the ER.

Main Methods:

  • Utilized the ER glycosylation machinery to map the integration of CFTR TM1 and TM2 within the ER membrane.
  • Assessed the effects of G85E and G91R mutations on CFTR's conformational stability and ER integration profiles.

Main Results:

  • The G85E mutation destabilizes the TM1 conformation, leading to temperature-insensitive ER accumulation of immature CFTR.
  • The G91R mutation alters the TM1 ER integration profile, causing temperature-dependent misfolding due to the introduction of a charged residue.
  • CF-causing mutations with similar predicted structural effects can induce disparate molecular perturbations.

Conclusions:

  • CFTR TM span integration and conformational stability are critical for proper protein folding and function.
  • Specific missense mutations in CFTR TM domains lead to distinct molecular defects, including altered ER integration and stability.
  • Understanding these mutation-specific mechanisms is crucial for developing targeted therapies for cystic fibrosis.

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