CFTR mutations altering CFTR fragmentation. / Tosoni, Kendra; Stobbart, Michelle; Cassidy, Diane M; Venerando, Andrea; Pagano, Mario A; Luz, Simao; Amaral, Margarida D; Kunzelmann, Karl; Pinna, Lorenzo A; Farinha, Carlos Miguel; Mehta, Anil.
In: Biochemical Journal, Vol. 449, No. 1, 2013, p. 295-305.Research output: Contribution to journal › Article
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TY - JOUR
T1 - CFTR mutations altering CFTR fragmentation
A1 - Tosoni,Kendra
A1 - Stobbart,Michelle
A1 - Cassidy,Diane M
A1 - Venerando,Andrea
A1 - Pagano,Mario A
A1 - Luz,Simao
A1 - Amaral,Margarida D
A1 - Kunzelmann,Karl
A1 - Pinna,Lorenzo A
A1 - Farinha,Carlos Miguel
A1 - Mehta,Anil
AU - Tosoni,Kendra
AU - Stobbart,Michelle
AU - Cassidy,Diane M
AU - Venerando,Andrea
AU - Pagano,Mario A
AU - Luz,Simao
AU - Amaral,Margarida D
AU - Kunzelmann,Karl
AU - Pinna,Lorenzo A
AU - Farinha,Carlos Miguel
AU - Mehta,Anil
PY - 2013
Y1 - 2013
N2 - Most cystic fibrosis (CF), results from deletion of a phenylalanine (F508) in the CF transmembrane-conductance regulator (CFTR, ABCC7) which causes ER-degradation of the mutant. Using stably CFTR-expressing BHK cell lines, we demonstrate that wild type- and F508delCFTR are cleaved into differently sized, N- and C-terminal bearing fragments, with each hemi-CFTR carrying its nearest nucleotide binding domain (NBD), reflecting differential cleavage through the central CFTR R-domain. Similar NBD1-bearing fragments are present in the natively expressing HBE epithelial cell line. We also observe multiple smaller fragments of different sizes in BHK cells, particularly after F508del-mutation (ladder pattern). Trapping wild type CFTR in the ER did not generate a F508del fragmentation fingerprint. Fragments change their size/pattern again post mutation at sites involved in CFTR's in vitro interaction with the pleiotropic protein kinase CK2 (S511A in NBD1). F508del- and S511A- mutation generate different fragmentation fingerprints that are each unlike wild type; yet both mutants generate a new N-terminal-bearing CFTR fragments that are not observed with other CK2-related mutations (S511D, S422A/D, T1471A/D). We conclude that F508delCFTR is not degraded completely and there exists a relationship between CFTR's fragmentation fingerprint and the CFTR sequence through putative CK2-interactive sites that lie near F508.
AB - Most cystic fibrosis (CF), results from deletion of a phenylalanine (F508) in the CF transmembrane-conductance regulator (CFTR, ABCC7) which causes ER-degradation of the mutant. Using stably CFTR-expressing BHK cell lines, we demonstrate that wild type- and F508delCFTR are cleaved into differently sized, N- and C-terminal bearing fragments, with each hemi-CFTR carrying its nearest nucleotide binding domain (NBD), reflecting differential cleavage through the central CFTR R-domain. Similar NBD1-bearing fragments are present in the natively expressing HBE epithelial cell line. We also observe multiple smaller fragments of different sizes in BHK cells, particularly after F508del-mutation (ladder pattern). Trapping wild type CFTR in the ER did not generate a F508del fragmentation fingerprint. Fragments change their size/pattern again post mutation at sites involved in CFTR's in vitro interaction with the pleiotropic protein kinase CK2 (S511A in NBD1). F508del- and S511A- mutation generate different fragmentation fingerprints that are each unlike wild type; yet both mutants generate a new N-terminal-bearing CFTR fragments that are not observed with other CK2-related mutations (S511D, S422A/D, T1471A/D). We conclude that F508delCFTR is not degraded completely and there exists a relationship between CFTR's fragmentation fingerprint and the CFTR sequence through putative CK2-interactive sites that lie near F508.
U2 - 10.1042/BJ20121240
DO - 10.1042/BJ20121240
M1 - Article
JO - Biochemical Journal
JF - Biochemical Journal
IS - 1
VL - 449
SP - 295
EP - 305
ER -