N-terminal amino acid sequence, immunohistochemical localization and tissue distribution of a plasma membrane protein (Prot17) of rat enterocytes.

Prot17, a protein of the basolateral membrane of rat small intestine with a mol.wt. of 17 kDa, can be isolated using a previously described method (Schiechl 1988). It occurs in the membrane as an oligomer with a mol.wt. of 90 kDa. In the present study a polyclonal antibody specific for Prot17 was used to explore by immunohistochemical techniques the tissue distribution of Prot17 and its ultrastructural localization within the cells. Furthermore the amino acid sequence of the N-terminal part of this molecule up to position 17 could be analyzed. The results are summarized as follows: Prot17 is a membrane anchored protein. Its partial amino acid sequence suggests that it is neither identical nor related to other known proteins. Immunofluorescence studies revealed, that it occurs only in epithelial cells. It is mainly found in the absorptive and goblet cells of the intestine and the acinar cells of the pancreas. Smaller quantities are found also in the bile duct epithelium of the liver, in the proximal tubule cells of the kidney and in the cells of the respiratory epithelium. Ultrastructural localization of Prot17 was possible in the intestinal epithelium and pancreas acinar cells. In both cell types it was found in the basolateral and microvillous membrane. In pancreas, Prot17 was also detected in the membrane of the zymogen granules. In the absorptive cells of the intestine Prot17 was found in both the membrane and the contents of subluminal vesicles. Furthermore, in apical granules of secretory cells of the respiratory epithelium binding of Prot17 specific antibody was found in the granular content, the membrane being negative.