In vivo antibody-mediated modulation of aminopeptidase A in mouse proximal tubular epithelial cells.
Aminopeptidase A (APA) is one of the many renal hydrolases. In mouse kidney, APA is predominantly expressed on the brush borders and sparsely on the basolateral membranes of proximal tubular epithelial cells. However, when large amounts of monoclonal antibodies (MAbs) against APA were injected into mice, we observed strong binding of the MAbs to the basolateral membranes, whereas the MAbs bound only transiently to the brush borders of the proximal tubular epithelial cells. In parallel, APA itself disappeared from the brush borders by both endocytosis and shedding, whereas it was increasingly expressed on the basolateral sides. Using ultrastructural immunohistology, we found no evidence for transcellular transport of endocytosed APA to the basolateral side of the proximal tubular epithelial cells. The absence of transcellular transport was confirmed by experiments in which we used a low dose of the MAbs. Such a low dose did not result in binding of the MAbs to the brush borders and had no effect on the presence of APA in the brush borders of the proximal tubular epithelial cells. In these experiments we still could observe binding of the MAbs to the basolateral membranes in parallel with the local appearance of APA. In addition, treatment of mice with chlorpromazine, a calmodulin antagonist that interferes with cytoskeletal function, largely inhibited the MAb-induced modulation of APA. Our studies suggest that injection of MAbs to APA specifically interrupts the normal intracellular traffic of this enzyme in proximal tubular epithelial cells. This intracellular transport is dependent on the action of cytoskeletal proteins.
Mentzel
S
S
Departments of Pathology, Division of Nephrology, University Hospital Nijmegen, Nijmegen, The Netherlands.
Dijkman
H B
HB
van Son
J P
JP
Wetzels
J F
JF
Assmann
K J
KJ
D000626:Aminopeptidases / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002746:Chlorpromazine / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D043384:Glutamyl Aminopeptidase; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000626:Aminopeptidases; D043384:Glutamyl Aminopeptidase; D008666:Metalloendopeptidases; D002746:Chlorpromazine