Identification and localization of a dogfish homolog of human cystic fibrosis transmembrane conductance regulator.
Chloride channels in the apical plasma membrane of cells in the dogfish rectal gland have served as a model system for the study of regulation of chloride flux by changes in intracellular cyclic AMP levels. Similar regulation by cyclic AMP has been described for channels in cells of human secretory epithelia where defective regulation by cyclic AMP-dependent protein phosphorylation is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). We have isolated a cDNA clone from the rectal gland encoding a protein that is 72% identical to the human CFTR. One of the major phosphorylation sites in CFTR is absent in the dogfish protein. The dogfish protein has, however, four additional putative substrate sites for the cyclic AMP-dependent protein kinase. A peptide antibody, which was raised against an amino acid sequence common to both the human and dogfish CFTR sequences, recognizes proteins with similar molecular masses (160 kDa) in the dogfish gland and in mammalian lung. Immunolocalization studies with this antibody show that the putative dogfish CFTR is localized to the apical membrane of cells lining the lumen of the rectal gland.
Marshall
J
J
Department of Pharmacology, School of Medicine, Yale University, New Haven, Connecticut 06510.
Martin
K A
KA
Picciotto
M
M
Hockfield
S
S
Nairn
A C
AC
Kaczmarek
L K
LK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004284:Dogfish / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011494:Protein Kinases / Q000378:metabolism; D012007:Rectum / Q000502:physiology; D012627:Sebaceous Glands / Q000166:cytology / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
C505032:CFTR protein, human; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA; D011494:Protein Kinases