Immunochemical identity of dipeptidyl aminopeptidase IV from pig serum, liver, submaxillary gland and kidney.

The enzymes which were extracted by autodigestion from the microsomal fractions of the pig kidney, liver and submaxillary gland and from the serum showed an immunochemical identity by a double immunodiffusion test. But the kidney enzyme had a different pI-value from the pI-values of the enzymes of other organs.     

Characterization of a soluble form of dipeptidyl peptidase IV from pig liver

Soluble dipeptidyl peptidase IV (EC 3.4.14.5) was purified from the 100,000 X g supernatant fraction of pig liver homogenate. The purified enzyme had the same properties as, and immunological identity with, the membrane-bound enzyme which was described previously. However, the purified enzyme had a pattern of molecular heterogeneity different from the membrane-bound enzyme; this was shown by isoelectric focusing. Carbohydrate analysis revealed that the soluble enzyme contained glucose, which is not found in the membrane-bound one, and less fucose, mannose, and sialic acid than the latter. From these results, we conclude that the soluble form of dipeptidyl peptidase IV in pig liver is closely related to the membrane-bound enzyme, but is not simply a proteolytically solubilized product of it.     

Characterization of a soluble form of dipeptidyl peptidase IV from pig liver

Summary Soluble dipeptidyl peptidase IV (EC 3.4.14.5) was purified from the 100,000×g supernatant fraction of pig liver homogenate. The purified enzyme had the same properties as, and immunological identity with, the membrane-bound enzyme which was described previously. However, the purified enzyme had a pattern of molecular heterogeneity different from the membrane-bound enzyme; this was shown by isoelectric focusing. Carbohydrate analysis revealed that the soluble enzyme contained glucose, which is not found in the membrane-bound one, and less fucose, mannose, and sialic acid than the latter. From these results, we conclude that the soluble form of dipeptidyl peptidase IV in pig liver is closley related to the membrane-bound enzyme, but is not simply a proteolytically solubilized product of it.We would like to thank Miss S. Fukushima for technical assistance.     

Deficiency of membrane-bound dipeptidyl aminopeptidase IV in a certain rat strain

Summary The activity of membrane-bound dipeptidyl aminopeptidase IV (EC 3.4.14.5) was found to be markedly reduced in the Fischer 344 rat strain compared with that in the Wistar strain. Analysis of membrane proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Ouchterlony immunodiffusion revealed the specific absence of this enzyme molecule in the Fischer 344 strain.     

Deficiency of membrane-bound dipeptidyl aminopeptidase IV in a certain rat strain

The activity of membrane-bound dipeptidyl aminopeptidase IV (EC 3.4.14.5) was found to be markedly reduced in the Fischer 344 rat strain compared with that in the Wistar strain. Analysis of membrane proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Ouchterlony immunodiffusion revealed the specific absence of this enzyme molecule in the Fischer 344 strain.     

Denervation and reinnervation of the rat's submaxillary gland

Zusammenfassung Parasympathische chirurgische Denervierung der Submaxillarisdrüse der Ratte führt zu einer Sensibilisierung der Effektorzellen, einer reduzierten Gesamtaktivität der Cholin-Acetyltransferase innerhalb von 1–2 Tagen und zu einer Gewichtsabnahme der Drüsen innerhalb von 4 Tagen. Das erste Zeichen einer Reinnervation ist eine verminderte Sensibilisierung und eine gesteigerte Enzymaktivität nach etwa 2 Monaten; das Gewicht der denervierten Drüsen nimmt nach 5 Monaten zu. Die Reinnervation ist nach 10 Monaten noch nicht vollständig.     

Glutaminase I activity in guinea pig and rabbit liver and kidney tissues

Zusammenfassung Aktive Glutaminase I wurde in Meerschweinchen- und Kaninchenleber, in Nierengewebsextrakten und in Mitochondrienpräparaten der erwähnten Organe bei pH 7,2 nachgewiesen.dl-Glutaminsäure bewirkte eine ausgesprochene Hemmung des Enzyms nach Zusatz von Phosphat. In hypertonischer Sukrosenährlösung lässt sich eine Aktivierung durch Phosphatzusatz gewöhnlich nicht nachweisen, selbst nicht beim pH-Optimum 7,2, es sei denn bei sehr hohen Phosphatkonzentrationen.     

Increased activity of dipeptidyl peptidase IV in serum of hepatoma-bearing rats coincides with the loss of the enzyme from the hepatoma plasma membrane

The specific activity of dipeptidyl peptidase IV (DPP IV E.C. 3.4.14.-) in the plasma membrane of Morris hepatoma 9121 or hepatoma 7777 was 3.5% and 2.9%, respectively, of that in the plasma membrane of rat liver. The enzyme activity in the serum of hepatoma-bearing rats was 141% (hepatoma 91219) and 162% (hepatoma 7777) of the normal value. Cytochemical investigation showed that the DPP IV activity was almost completely absent from the hepatoma cell plasma membrane and was not sequestered within these cells. Indirect immunofluorescence staining with a polyclonal antibody directed against DPP IV indicated that the loss of activity was due to the absence of DPP IV molecules in the plasma membrane. The possibility that the enzyme is transferred from the membrane into the serum as a result of structural alterations is discussed.     

Increased activity of dipeptidyl peptidase IV in serum of hepatoma-bearing rats coincides with the loss of the enzyme from the hepatoma plasma membrane

Summary The specific activity of dipeptidyl peptidase IV (DPPIV E.C. 3.4.14.-) in the plasma membrane of Morris hepatoma 9121 or hepatoma 7777 was 3.5% and 2.9%, respectively, of that in the plasma membrane of rat liver. The enzyme activity in the serum of hepatoma-bearing rats was 141% (hepatoma 91219) and 162% (hepatoma 7777) of the normal value. cytochemical investigation showed that the DPP IV activity was almost completely absent from the hepatoma cell plasma membrane and was not sequestered within these cells. Indirect immunofluorescence staining with a polyclonal antibody directed against DPP IV indicated that the loss of activity was due to the absence of DPP IV molecules in the plasma membrane. The possibility that the enzyme is transferred from the membrane into the serum as a result of structural alterations is discussed.     

Degeneration activity after sympathetic denervation of the submaxillary gland and the eye

Zusammenfassung Nach Sympathectomie erscheint eine «Degenerationskontraktion» der Nickhaut nicht nur in wachen oder spinalen, sondern auch regelmässig in narkotisierten Katzen. Die «Degenerationssekretion» von der Submaxillarisdrüse tritt später auf; aber auf Grund der Variabilität der sympathisch sekretorischen Innervation sieht man sie nicht bei allen Tieren.

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Supported by a grant from the Swedish Medical Research Council.     

Dissociation-constants of metal-ion-complexes with alkaline phosphatase from pig kidney

Summary Using metal-ion buffers it was possible to remove Zn²⁺, Mg²⁺ and Mn²⁺ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are K_EMg:4·10^–7 M, K_EMn:4·10^–8 M and K_EZn:8·10^–13 M (22°C, pH:9.6, :0.07).Acknowledgement: The authors thank Dr.H. U. Wolf for helpful suggestions and valuable discussion and MissH. Köth for technical assistance.     

Analysis of vasodilatation in the submaxillary gland using potentiators of acetylcholine and kinins

Zusammenfassung Nachweis, dass bei Reizung des N. chorda lingualis der Katze ein Blutdruckeffekt ausbleibt, wenn ein Bradykinin potenzierender Faktor (Präparat BPF) zuvor gegeben wurde.

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Acknowledgments. We are greatly indepted to Dr.J. Stewart of the University of Colorado and to Dr.L. Greene of Brookhaven National Laboratories, New York, for kind gifts of synthetic bradykinin potentiators.

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This work was supported by grants from the Medical Research Council of Canada and the Alberta Heart Foundation.     

Secretory responses and choline acetylase of the rat's submaxillary gland after duct ligation

Zusammenfassung Abbinden des Ausführungsganges der Submaxillarisdrüse der Ratte führt zu ihrer Gewichtsabnahme. Nach i.v. Gaben von Methacholin und Adrenalin verschwindet die Sekretion innerhalb von 4–6 Tagen nach der Operation, während bei Pilokarpin noch eine sehr verminderte Speichelsekretion persistiert. Die spezifische Fähigkeit der sekretorischen Zellen war zusehends verringert. Die Gesamtaktivität der Cholin-Acetylase blieb ähnlich derjenigen der Kontrolldrüsen, was für eine unveränderte Funktion des Parasympathikus spricht.     

Dissociation-constants of metat-ion-complexes with alkaline phosphatase from pig kidney.

Using metal-ion buffers it was possible to remove Zn2+, Mg2+ and Mn2+ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are KEMg: 4 X 10(-7) M, KEMn: 4 X 10(-8) M and KEZn: 8 X 10(-13) M (22 degrees C, pH: 9.6, mu: 0.07).     

Effect of furosemide on aminopeptidase activity in the rat kidney and urine

Zusammenfassung Histochemische und biochemische Untersuchungen des Verhaltens von Aminopeptidasen nach Furosemidgabe zeigen einen Abfall bzw. Anstieg der Enzymaktivitäten in Nierenschnitten und im Urin von männlichen Wistar-Ratten. Die Resultate weisen auf eine direkte, Furosemid-induzierte reversible Epithelschädigung im proximalen Tubulus hin. Unter gleichen Bedingungen sind entsprechende Befunde bei weiblichen Wistar-Ratten nicht zu erheben.

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Supported by Deutsche Forschungsgemeinschaft.