Isoforms of the Lutheran/basal cell adhesion molecule glycoprotein are differentially delivered in polarized epithelial cells. Mapping of the basolateral sorting signal to a cytoplasmic di-leucine motif.
Lu and Lu(v13) are two glycoprotein (gp) isoforms that belong to the immunoglobulin superfamily and carry both the Lutheran (Lu) blood group antigens and the basal cell adhesion molecule epithelial cancer antigen. Lu (85 kDa) and Lu(v13) (78 kDa) gps, which differ only in the length of their cytoplasmic domain, are adhesion molecules that bind laminin. In nonerythroid tissues, the Lu/basal cell adhesion molecule antigens are predominantly expressed in the endothelium of blood vessel walls and in the basement membrane region of normal epithelial cells, whereas they exhibit a nonpolarized expression in some epithelial cancers. Here, we analyzed the polarization of Lu and Lu(v13) gps in epithelial cells by confocal microscopy and domain-selective biotinylation assays. Differentiated human colon carcinoma Caco-2 cells exhibited a polarized expression of endogenous Lu antigens associated with a predominant expression of the Lu isoform at the basolateral domain of the plasma membrane and a very low expression of the Lu(v13) isoform at both the apical and basolateral domains. Analysis of transfected Madin-Darby canine kidney cells revealed a basolateral expression of Lu gp and a nonpolarized expression of Lu(v13) gp. Delivery of Lu(v13) to both apical and basolateral surfaces showed similar kinetics, indicating that this isoform is directly transported to each surface domain. A dileucine motif at position 608-609, specific to the Lu isoform, was characterized as a dominant basolateral sorting signal that prevents Lu gp from taking the apical delivery pathway.
El Nemer
W
W
INSERM U76, Institut National de la Transfusion Sanguine, 6 rue Alexandre Cabanel, 75015 Paris, France.
Colin
Y
Y
Bauvy
C
C
Codogno
P
P
Fraser
R H
RH
Cartron
J P
JP
Le Van Kim
C L
CL
D000595:Amino Acid Sequence; D000818:Animals; D018938:Caco-2 Cells; D015815:Cell Adhesion Molecules / Q000502:physiology*; D016764:Cell Polarity; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism*; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008188:Lutheran Blood-Group System; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000502:physiology*; D010449:Peptide Mapping; D015398:Signal Transduction; D013329:Structure-Activity Relationship
C090228:BCAM protein, human; D015815:Cell Adhesion Molecules; D008188:Lutheran Blood-Group System; D009363:Neoplasm Proteins; D007930:Leucine