Suppressing role of transforming growth factor-beta 1 on cathepsin activity in cultured kidney tubule cells.
Elevated expression and activity of transforming growth factor-beta 1 (TGF-beta 1) have been indicated in various renal diseases, implicating the potential involvement of this growth factor in the accumulation of extracellular matrix. To assess its potential role on protein turnover in tubule cells, we investigated in LLC-PK1 cells the effects of TGF-beta 1 on the activities of lysosomal cysteine proteinases cathepsins B, H, and L + B, which play a major role in the degradation of both cellular protein and extracellular matrix. The results show that 1-10 ng/ml TGF-beta 1 exerted inhibitory effects on cathepsin B and L + B activities, when applied to either the basolateral or apical membrane of these cells (basolateral side: B -23.2%, L + B -19.9%; apical side: B -28.2%, L + B -22.6%). Application of TGF-beta 1 to both sides enhanced suppression of the enzyme activities (B -37.8%, L + B -37.4%). This suppression of cathepsin activities was accompanied by a reduction of cellular protein degradation rate by 20.0% after 24 h and 51.7% after 48 h. Furthermore, TGF-beta 1 stimulated cellular protein synthesis by 50.0% after 48 h. The combined effects on protein turnover resulted in cellular hypertrophy: increases of both protein content and cell size after 48 h. Concerning the underlying mechanism, TGF-beta 1 did not induce a rise in intracellular Ca2+ concentration nor did Ca2+ channel blocker verapamil (10(-6) M) ameliorate the TGF-beta 1-induced inhibition of cathepsin activities. However, TGF-beta 1 raised the pH in lysosomes, which obviously impaired the acidic cysteine proteinases. In conclusion, the TGF-beta 1-induced cellular hypertrophy is caused by both enhanced protein synthesis and reduced protein breakdown. Suppression of cathepsin B and L + B activities mediated probably by an alkalinization in lysosomes is involved in the decreased protein degradation.
Ling
H
H
Division of Nephrology, University of Würzburg, Germany.
Vamvakas
S
S
Busch
G
G
Dämmrich
J
J
Schramm
L
L
Lang
F
F
Heidland
A
A
D000818:Animals; D002021:Buffers; D002403:Cathepsins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002478:Cells, Cultured; D006863:Hydrogen-Ion Concentration; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism*; D018374:LLC-PK1 Cells; D011506:Proteins / Q000378:metabolism; D013552:Swine; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D002021:Buffers; D011506:Proteins; D016212:Transforming Growth Factor beta; D002403:Cathepsins