GP2, a GPI-anchored protein in the apical plasma membrane of the pancreatic acinar cell, co-immunoprecipitates with src kinases and caveolin.
We previously showed that endocytosis at the apical plasma membrane (APM) of the pancreatic acinar cell is activated by the cleavage of GP2, a GPI-linked protein, from the apical cell surface. This endocytic process, as measured by horseradish peroxidase uptake into pancreatic acinar cells, is blocked by the tyrosine kinase inhibitors genistein and tyrphostin B42 as well as by disruption of actin filaments with cytochalasin. This suggests that the cleavage of GP2 from the cell membrane may activate endocytosis through a tyrosine kinase-regulated pathway. However, the mechanism by which GP2 and tyrosine kinases act together to activate endocytosis at the APM remains unknown. In this study, we demonstrate that pp60, p62yes, caveolin, and annexin, which have previously been implicated in endocytosis in other cell lines, were present in high abundance in GPI-enriched membranes by Western blot analysis. pp60, p62yes, and caveolin all co-immunoprecipitated with GP2 except annexin. An 85-kDa protein whose tyrosine-dependent phosphorylation is correlated with the activation of endocytosis in intact acinar cells also was present in these immunoprecipitates. This suggests that in pancreatic acini, GP2 may exist in a complex with src kinases, caveolin, and an 85-kDa phosphorylated substrate to regulate endocytosis at the APM.
Parker
E M
EM
The Pancreas Center, Beth Israel Deaconess Medical Center, Harvard Medical Schlool, Boston, Massachusetts 02215, USA.
Zaman
M M
MM
Freedman
S D
SD
D000818:Animals; D017302:Annexins / Q000032:analysis; D015153:Blotting, Western; D051242:Caveolin 1; D022461:Caveolins / Q000032:analysis*; D002462:Cell Membrane / Q000737:chemistry*; D004705:Endocytosis; D058851:GPI-Linked Proteins; D007163:Immunosorbent Techniques; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D010179:Pancreas / Q000737:chemistry*; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000032:analysis; D051575:Proto-Oncogene Proteins c-yes; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000032:analysis; D019061:src-Family Kinases / Q000032:analysis*
D017302:Annexins; C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; C489339:GP2 protein, human; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D011518:Proto-Oncogene Proteins; D051575:Proto-Oncogene Proteins c-yes; D016392:Proto-Oncogene Proteins pp60(c-src); C496639:Yes1 protein, mouse; D019061:src-Family Kinases