Human CD8 alpha glycoprotein is expressed at the apical plasma membrane domain in permanently transformed MDCK II clones.
Madin-Darby canine kidney cells (MDCK II) have been cotransfected with plasmids expressing the human CD8 alpha glycoprotein and the bacterial gene which confers resistance to neomycin. Stable transformants have been isolated in the presence of G-418 in the culture medium and screened for CD8 alpha expression by immunofluorescence. The three clones we have characterized showed: 1) high level of synthesis and efficient surface expression of glycosylated, homodimeric CD8 alpha and 2) preferential apical deposition of CD8 alpha in confluent monolayers. This polar distribution has been measured in cells grown on a plastic substratum as well as on nitrocellulose filter by means of EM immunocytochemistry and surface radioimmunoassay. CD8 alpha was 6 to 11-fold enriched on the apical membrane whereas the 58 kDa protein, a basolateral marker in MDCK II cells, resulted about 9-fold enriched on the basolateral membrane of the three clones. We believe these permanently transformed clones could prove to be a useful tool with which to study cell polarity.
Migliaccio
G
G
Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli, Italy.
Zurzolo
C
C
Nitsch
L
L
Obici
S
S
Lotti
L V
LV
Torrisi
M R
MR
Pascale
M C
MC
Leone
A
A
Bonatti
S
S
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis*; D016827:CD8 Antigens; D002461:Cell Line, Transformed / Q000378:metabolism* / Q000648:ultrastructure; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002999:Clone Cells; D004285:Dogs; D005622:Friend murine leukemia virus / Q000235:genetics; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D010969:Plastics; D011863:Radioimmunoassay; D013462:Sulfur Radioisotopes; D013997:Time Factors; D014162:Transfection / Q000235:genetics
D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D008562:Membrane Glycoproteins; D010969:Plastics; D013462:Sulfur Radioisotopes