TY - JOUR
T1 - CFTR Folding Consortium
T2 - methods available for studies of CFTR folding and correction.
AU - Peters, Kathryn W.
AU - Okiyoneda, Tsukasa
AU - Balch, William E.
AU - Braakman, Ineke
AU - Brodsky, Jeffrey L.
AU - Guggino, William B.
AU - Penland, Christopher M.
AU - Pollard, Harvey B.
AU - Sorscher, Eric J.
AU - Skach, William R.
AU - Thomas, Philip J.
AU - Lukacs, Gergely L.
AU - Frizzell, Raymond A.
PY - 2011
Y1 - 2011
N2 - The CFTR Folding Consortium (CFC) was formed in 2004 under the auspices of the Cystic Fibrosis Foundation and its drug discovery and development affiliate, CFF Therapeutics. A primary goal of the CFC is the development and distribution of reagents and assay methods designed to better understand the mechanistic basis of mutant CFTR misfolding and to identify targets whose manipulation may correct CFTR folding defects. As such, reagents available from the CFC primarily target wild-type CFTR NBD1 and its common variant, F508del, and they include antibodies, cell lines, constructs, and proteins. These reagents are summarized here, and two protocols are described for the detection of cell surface CFTR: (a) an assay of the density of expressed HA-tagged CFTR by ELISA and (b) the generation and use of an antibody to CFTR's first extracellular loop for the detection of endogenous CFTR. Finally, we highlight a systematic collection of assays, the CFC Roadmap, which is being used to assess the cellular locus and mechanism of mutant CFTR correction. The Roadmap queries CFTR structure-function relations at levels ranging from purified protein to well-differentiated human airway primary cultures.
AB - The CFTR Folding Consortium (CFC) was formed in 2004 under the auspices of the Cystic Fibrosis Foundation and its drug discovery and development affiliate, CFF Therapeutics. A primary goal of the CFC is the development and distribution of reagents and assay methods designed to better understand the mechanistic basis of mutant CFTR misfolding and to identify targets whose manipulation may correct CFTR folding defects. As such, reagents available from the CFC primarily target wild-type CFTR NBD1 and its common variant, F508del, and they include antibodies, cell lines, constructs, and proteins. These reagents are summarized here, and two protocols are described for the detection of cell surface CFTR: (a) an assay of the density of expressed HA-tagged CFTR by ELISA and (b) the generation and use of an antibody to CFTR's first extracellular loop for the detection of endogenous CFTR. Finally, we highlight a systematic collection of assays, the CFC Roadmap, which is being used to assess the cellular locus and mechanism of mutant CFTR correction. The Roadmap queries CFTR structure-function relations at levels ranging from purified protein to well-differentiated human airway primary cultures.
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U2 - 10.1007/978-1-61779-120-8_20
DO - 10.1007/978-1-61779-120-8_20
M3 - Article
C2 - 21547742
AN - SCOPUS:80052541318
SN - 1064-3745
VL - 742
SP - 335
EP - 353
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -