Studies on human porin. IX. Immunolocalization of porin and CFTR channels in human surface respiratory epithelium.
The expression of the voltage-dependent anion channel (VDAC) "Porin 31 HL" and its cellular and subcellular immunocytochemical localization in the human respiratory epithelium were studied with monoclonal and polyclonal antibodies using immunofluorescence and immunogold labelling with light (LM) and transmission electron microscopy (TEM), respectively. Porin was identified in the apical domain of the ciliated cells and in the basal cells of the respiratory epithelium. Immunogold labelling was present in the apical plasma membrane and subapical vesicles of the ciliated cells. In pre-embedded freshly dissociated surface epithelial cells, porin could also be identified with TEM at the outer part of the plasma membrane of basal cells. By LM double immunolabelling, both porin and cystic fibrosis transmembrane conductance regulator (CFTR) were identified in the apical domain of ciliated cells but not in basal cells where CFTR was never identified. On Western blots of solubilized total membrane protein preparations from the same frozen surface epithelial respiratory cells, the antibodies recognized a group of 3 proteins of 31, 60 and 130-140 kDa with a strong reactivity for a 31 kDa protein, corresponding to the porin and a protein of 170 kDa which is consistent with mature CFTR. These results suggest that porin might be part of a multi-component chloride channel complex which could interact with CFTR.
Puchelle
E
E
Unité INSERM 314, CHR Maison Blanche, Université de Reims.
Jacquot
J
J
Fuchey
C
C
Burlet
H
H
Klossek
J M
JM
Gilain
L
L
Triglia
J M
JM
Thinnes
F P
FP
Hilschmann
N
N
D000906:Antibodies / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007473:Ion Channels / Q000378:metabolism* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008854:Microscopy, Electron; D018272:Porins; D012137:Respiratory System / Q000378:metabolism* / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000378:metabolism
D000906:Antibodies; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; C505032:CFTR protein, human; D007473:Ion Channels; D008565:Membrane Proteins; D018272:Porins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator