Identification of an 85-100 kDa glycoprotein as a cell surface marker for an advanced stage of urothelial differentiation: association with the inter-plaque ('hinge') area.
Although bladder cancers account for almost 5% of all human cancer deaths, little is known about the biochemistry of urothelial differentiation. We have recently identified three major protein subunits ('uroplakins') of asymmetric unit membranes (AUM), which form rigid-looking plaques covering up to 70% of the apical surface of urothelial superficial (umbrella) cells. The ordinary-looking plasma membranes that interconnect these plaques are believed to be functionally specialized, serving as flexible but durable 'hinges'. Whether these hinge membranes are biochemically unique is unknown. Using a new monoclonal antibody (AE32) we have identified an 85-100 kDa glycoprotein (UGP85) which appears to be urothelium-specific. In both normal urothelium and cultured urothelial colonies this cell surface protein is associated mainly with superficial cells, suggesting that its expression is differentiation dependent. Results from in vitro translation experiments indicated that this glycoprotein contains a core polypeptide of about 55 kDa. Using immunogold localization techniques, we showed that in cultured urothelial colonies--which are known to lack mature AUM plaques--UGP85 is distributed relatively uniformly on the apical surface of some differentiated cells. However, in superficial cells of normal urothelium UGP85 is mainly associated with the hinge areas. These results raise the possibility that UGP85 is a plasma membrane component which can be excluded, to varying extents, from the plaque region as 12 nm protein particles are assembled into a tightly packed paracrystalline AUM structure. The identification of UGP85 provides the first evidence that the hinge areas interconnecting the urothelial plaques are biochemically distinguishable from the plasma membranes of the relatively undifferentiated urothelial cells of the lower cell layers.(ABSTRACT TRUNCATED AT 250 WORDS)
Yu
J
J
Department of Dermatology, New York University Medical School, New York 10016.
Manabe
M
M
Sun
T T
TT
D000818:Animals; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002417:Cattle; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006031:Glycosylation; D007150:Immunohistochemistry; D008970:Molecular Weight; D001743:Urinary Bladder / Q000166:cytology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D006023:Glycoproteins