Abstract

The working hypothesis driving this proposal is that there are one or two anion binding sites within the pore and that the best probes of the pore are permeant anions themselves. Experimental methods combine expression of CFTR in Xenopus oocytes with single channel and macroscopic recording. In some cases, L-cells which stably express CFTR may be used (these cells will be provided by Dr. R. Bridges). Single channel recording is critical to confirm changes in conductance and mole fraction effects while permeation ratios can be easily determined from macroscopic currents. Controls will be in the form of non-CFTR expressing oocytes and oocytes expressing CFTR protein, but have not been stimulated to express current. The endogenous IClCa found in oocytes will be reduced by replacing extracellular Ca++ with Ba++. This proposal is focused on anion permeation independent of CFTR gating issues. These techniques will be used to develop a model of anion permeation for CFTR.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK045880-06
Application #
2394430
Study Section
Physiology Study Section (PHY)
Program Officer
Mckeon, Catherine T
Project Start
1992-09-30
Project End
2001-09-29
Budget Start
1997-09-30
Budget End
1998-09-29
Support Year
6
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of Michigan Ann Arbor
Department
Physiology
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109

  Publications

Moran, A R; Norimatsu, Y; Dawson, D C et al. (2014) Aqueous cigarette smoke extract induces a voltage-dependent inhibition of CFTR expressed in Xenopus oocytes. Am J Physiol Lung Cell Mol Physiol 306:L284-91
Liu, Xuehong; Dawson, David C (2014) Cystic fibrosis transmembrane conductance regulator (CFTR) potentiators protect G551D but not ?F508 CFTR from thermal instability. Biochemistry 53:5613-8
Norimatsu, Yohei; Ivetac, Anthony; Alexander, Christopher et al. (2012) Cystic fibrosis transmembrane conductance regulator: a molecular model defines the architecture of the anion conduction path and locates a ""bottleneck"" in the pore. Biochemistry 51:2199-212
Norimatsu, Yohei; Ivetac, Anthony; Alexander, Christopher et al. (2012) Locating a plausible binding site for an open-channel blocker, GlyH-101, in the pore of the cystic fibrosis transmembrane conductance regulator. Mol Pharmacol 82:1042-55
Liu, Xuehong; O'Donnell, Nicolette; Landstrom, Allison et al. (2012) Thermal instability of ýýF508 cystic fibrosis transmembrane conductance regulator (CFTR) channel function: protection by single suppressor mutations and inhibiting channel activity. Biochemistry 51:5113-24
Norimatsu, Yohei; Moran, Aurelia R; MacDonald, Kelvin D (2012) Lubiprostone activates CFTR, but not ClC-2, via the prostaglandin receptor (EP(4)). Biochem Biophys Res Commun 426:374-9
Liu, Xuehong; Dawson, David C (2011) Cystic fibrosis transmembrane conductance regulator: temperature-dependent cysteine reactivity suggests different stable conformers of the conduction pathway. Biochemistry 50:10311-7
Alexander, Christopher; Ivetac, Anthony; Liu, Xuehong et al. (2009) Cystic fibrosis transmembrane conductance regulator: using differential reactivity toward channel-permeant and channel-impermeant thiol-reactive probes to test a molecular model for the pore. Biochemistry 48:10078-88
Liu, Xuehong; Alexander, Christopher; Serrano, Jose et al. (2006) Variable reactivity of an engineered cysteine at position 338 in cystic fibrosis transmembrane conductance regulator reflects different chemical states of the thiol. J Biol Chem 281:8275-85
Weber, Gerhard J; Mehr, Ali Poyan; Sirota, Jeffrey C et al. (2006) Mercury and zinc differentially inhibit shark and human CFTR orthologues: involvement of shark cysteine 102. Am J Physiol Cell Physiol 290:C793-801

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