Calreticulin is transported to the surface of NG108-15 cells where it forms surface patches and is partially degraded in an acidic compartment.
Although calreticulin (Crt) is primarily localized to the endoplasmic reticulum (ER), our results using biotinylation and immunocytochemical methods indicate that a small but significant amount of Crt is present and forms large patches on the surface of NG108-15 cells (a mouse neuroblastoma-rat glioma hybrid cell line). (35)S-labelled Crt molecules begin to reach the cell surface after only 10 min of labelling and disappear slowly from the cell surface. After 4 hr of labelling, approximately half of the newly synthesized Crt molecules are on the cell surface. We believe that some Crt molecules may escape from the KDEL receptor-mediated salvage pathway as they are synthesized and proceed through the secretory pathway to the cell surface. Immunoprecipitation from the culture medium shows that Crt is not released from the cell surface to the medium, suggesting tight binding to surface molecules. NH(4)Cl can block the degradation of Crt; therefore, Crt is presumably degraded in the lysosome pathway. However, blockage of the disappearance of surface Crt is less efficient than that of internal Crt. This suggests that the disappearance of Crt from the cell surface may not be due solely to its degradation, but may reflect transport into another cell compartment such as the ER.
Xiao
G
G
Department of Chemistry, Boston University, Boston, Massachusetts, USA.
Chung
T F
TF
Fine
R E
RE
Johnson
R J
RJ
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D019921:Biotinylation; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D002451:Cell Compartmentation / Q000502:physiology*; D002462:Cell Membrane / Q000737:chemistry* / Q000502:physiology; D006822:Hybrid Cells / Q000378:metabolism*; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D011233:Precipitin Tests; D051381:Rats; D012261:Ribonucleoproteins / Q000378:metabolism*; D013997:Time Factors
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D008565:Membrane Proteins; D012261:Ribonucleoproteins