Expression of cobalamin transport proteins and cobalamin transcytosis by colon adenocarcinoma cells.
Human colon adenocarcinoma (Caco-2) cells express both intrinsic factor-cobalamin receptor and transcobalamin II (TC II). The expression of these activities began to rise by day 6 and reached peak levels between 10 and 15 days in culture. The postconfluent Caco-2 cell membranes bound approximately 30-35 fmol of intrinsic factor (IF) [57Co]Cbl/mg protein. The size of the mature receptor expressed in the apical brush border had a relative molecular mass of 230 kDa. The intracellular form of TC II had a Mr of 43, 5 higher than the secreted form of TC II. TC II was secreted unidirectionally via the basolateral direction when Caco-2 cells were grown on culture inserts. When grown on culture inserts, the Caco-2 cells were polarized (electrical resistance greater than 200 omega/cm2) and transcytosed [57Co]Cbl bound to IF from apical-to-basal but not from basal-to-apical direction. Under these conditions, [57Co]Cbl complexed to haptocorrin was not transported. These cells also transcytosed free [57Co]Cbl, although less efficiently. The [57Co]Cbl transcytosed using either IF[57Co]Cbl or free [57Co]Cbl as ligands was bound exclusively to TC II. Intracellular [57Co]Cbl decreased during transcytosis with a slow (t1/2 = 4 h) transfer of [57Co]Cbl from IF to TC II. These results show that the transport of Cbl in Caco-2 cells is very similar to the human enterocyte system.
Ramanujam
K S
KS
Department of Medicine, Medical College of Wisconsin, Milwaukee 53226.
Seetharam
S
S
Ramasamy
M
M
Seetharam
B
B
D000230:Adenocarcinoma; D001692:Biological Transport; D002460:Cell Line; D003110:Colonic Neoplasms; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007437:Intrinsic Factor / Q000096:biosynthesis* / Q000302:isolation & purification; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D013462:Sulfur Radioisotopes; D014155:Transcobalamins / Q000096:biosynthesis* / Q000302:isolation & purification; D014805:Vitamin B 12 / Q000378:metabolism
D013462:Sulfur Radioisotopes; D014155:Transcobalamins; D007437:Intrinsic Factor; D008715:Methionine; D014805:Vitamin B 12