Cyclosporin inhibits phosphate transport and stimulates alkaline phosphatase activity in renal BBMV.
The activity of various enzymes and transport systems was studied in renal brush-border membrane vesicles (BBMV) isolated from rats injected daily with cyclosporin. Alkaline phosphatase (AP) was strongly stimulated: 55 and 113% increases were obtained in BBMV isolated from rats injected with 10 mg cyclosporin/kg for 5 and 10 days. The affinity of the enzyme remained unaltered, but maximal activity (Vmax) showed a strong increase of 2.4-fold between control and treated animals. In addition to the phosphatase activity, phosphate binding to AP also showed a dose-dependent stimulation by cyclosporin treatment: 44 and 70% increases in animals treated for 5 days with 5 and 10 mg cyclosporin/kg. However, the activity of aminopeptidase M was not affected by these treatments, and polyacrylamide gel electrophoresis of BBMV revealed no alterations in the profile of membrane proteins, suggesting the specificity of cyclosporin interaction with alkaline phosphatase. Na(+)-dependent amino acid and D-glucose transport systems remained unaffected by cyclosporin treatment. The Na(+)-independent transport system for lysine and the Na(+)-H+ antiporter activity were also unaltered. In contrast, the initial rate of phosphate uptake decreased by 28% after administration of cyclosporin (10 mg/kg) for 5 days: the Michaelis constant (Km) and Vmax decreased from 137 to 85 microM and from 1.49 to 1.07 pmol.micrograms-1.5 s-1, respectively. "In vitro" studies with membranes isolated from untreated rats were also undertaken by preincubating membranes with cyclosporin. Neither alkaline phosphatase nor the transport systems were affected under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)
Demeule
M
M
Département de Chimie, Université du Québec à Montréal, Canada.
Béliveau
R
R
D000042:Absorption; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D018826:CD13 Antigens; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003524:Cyclosporins / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D008871:Microvilli / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism* / Q000652:urine; D051381:Rats; D011919:Rats, Inbred Strains; D012964:Sodium / Q000494:pharmacology
D003524:Cyclosporins; D008565:Membrane Proteins; D010710:Phosphates; D012964:Sodium; D000469:Alkaline Phosphatase; D000626:Aminopeptidases; D018826:CD13 Antigens