Intracrystalline proteins and calcium oxalate crystal degradation in MDCK II cells.
We assessed the effects of intracrystalline urinary proteins on the ability of Type II Madin-Darby canine kidney (MDCK-II) cells to bind and degrade calcium oxalate monohydrate (COM) crystals. Binding of [14C]-labelled inorganic crystals (iCOM), and COM crystals precipitated from centrifuged and filtered (CF) or ultrafiltered (UF) human urine was quantified by radioactive analysis. SDS-PAGE confirmed the presence of intracrystalline proteins > 10 kDa in CF crystals and their absence from UF crystals. Morphological effects were assessed qualitatively by field emission scanning electron microscopy. iCOM crystals bound rapidly and extensively and were resistant to degradation. Binding of CF crystals was weaker than UF crystals, and both had markedly less affinity than iCOM. CF and UF crystals were extensively degraded within 90 min, the effect being more pronounced with CF. These results support our hypothesis that intracrystalline proteins protect against urolithiasis by facilitating intracellular proteolytic digestion and destruction of crystals phagocytosed by urothelial cells.
Chauvet
Magali C
MC
Department of Surgery, Flinders Medical Centre and Flinders University School of Medicine, Bedford Park, SA 5042, Australia. magali.chauvet@flinders.edu.au
Ryall
Rosemary Lyons
RL
D000818:Animals; D001711:Biotransformation / Q000187:drug effects; D002129:Calcium Oxalate / Q000737:chemistry / Q000378:metabolism* / Q000652:urine; D002478:Cells, Cultured; D003460:Crystallization; D004285:Dogs; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D014556:Urine / Q000737:chemistry
D011506:Proteins; D002129:Calcium Oxalate