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| PURIFICATION AND FUNCTIONAL RECONSTITUTION OF THE CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR (CFTR) |
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| Author(s): BEAR CE, LI CH, KARTNER N, BRIDGES RJ, JENSEN TJ, RAMJEESINGH M, RIORDAN JR |
| Source: CELL Volume: 68 Issue: 4 Pages: 809-818 Published: FEB 21 1992 |
| Times Cited: 649 References: 44 |
| Abstract: Circumstantial evidence has accumulated suggesting that CFTR is a regulated low-conductance CI- channel. To test this postulate directly, we have purified to homogeneity a recombinant CFTR protein from a high-level baculovirus-infected insect cell line. Evidence of purity included one- and two-dimensional gel electrophoresis, N-terminal peptide sequence, and quantitative amino acid analysis. Reconstitution into proteoliposomes at less than one molecule per vesicle was accomplished by established procedures. Nystatin and ergosterol were included in these vesicles, so that nystatin conductance could serve as a quantitative marker of vesicle fusion with a planar lipid bilayer. Upon incorporation, purified CFTR exhibited regulated chloride channel activity, providing evidence that the protein itself is the channel. This activity exhibited the basic biophysical and regulatory properties of the type of CI- channel found exclusively in CFTR-expressing cell types and believed to underlie cAMP-evoked secretion in epithelial cells. |
| Document Type: Article |
| Language: English |
| Reprint Address: BEAR, CE (reprint author), UNIV TORONTO, DEPT PHYSIOL, TORONTO M5S 1A8, ONTARIO CANADA |
Addresses:
1. UNIV TORONTO, DEPT BIOCHEM, TORONTO M5S 1A8, ONTARIO CANADA
2. UNIV TORONTO, DEPT CLIN BIOCHEM, TORONTO M5S 1A8, ONTARIO CANADA
3. HOSP SICK CHILDREN, RES INST, TORONTO M5G 1X8, ONTARIO CANADA
4. UNIV ALABAMA, DEPT PHYSIOL & BIOPHYS, BIRMINGHAM, AL 35294 USA |
| Publisher: CELL PRESS, 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 |
| Subject Category: Biochemistry & Molecular Biology; Cell Biology |
| IDS Number: HF440 |
| ISSN: 0092-8674 |
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