Biochemical properties of dipeptidyl peptidase IV in liver and hepatoma plasma membranes.
In the present investigation we compared the glycoprotein DPP IV from rat liver and Morris hepatoma 7777 by means of biochemical and immunological methods. For that purpose nine monoclonal anti-DPP IV-antibodies recognizing four different epitopes and a monospecific anti-DPP IV-antiserum were applied. In the homogenates of both tissues a plasma membrane-bound and a soluble form were detected. The immunological cross-reactivity of both forms was demonstrated with the antiserum and the monoclonal antibodies against the epitopes A, B and C while epitope D was restricted to liver plasma membrane. Differences of the distinct DPP IV forms were exhibited in the molecular weights, isoelectric points and peptide maps. In the hepatoma homogenate only 10% of DPP IV activity was found compared to normal liver but the ratio of soluble to membrane-bound form is higher in the hepatoma than in the liver. The fractionation of the homogenates into different cell components revealed for the liver a continuous increase of DPP IV activity from the endoplasmic reticulum fractions to the Golgi apparatus and finally to the plasma membranes. By contrast, in hepatoma the flow from the Golgi apparatus to plasma membrane was greatly reduced. The loss of DPP IV from the surface of cultured hepatoma cells was concomitant with a decrease of cell-substratum adhesion. DPP IV was found to be inserted into the liver plasma membrane by two different mechanisms, a phospholipase C-sensitive and a papain-sensitive one. In the hepatoma the phospholipase C-sensitive anchorage was not expressed. Besides liver and hepatoma the distribution of DPP IV was characterized in various rat organs by enzyme activity, histochemistry and immunohistochemistry with the anti-DPP IV-antibodies.
Reutter
W
W
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, F.R.G.
Hartel
S
S
Hanski
C
C
Huhle
T
T
Zimmer
T
T
Gossrau
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism*; D015195:Drug Design; D000939:Epitopes / Q000032:analysis; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008099:Liver / Q000201:enzymology*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology*; D010449:Peptide Mapping; D051381:Rats; D014407:Tumor Cells, Cultured / Q000201:enzymology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4