Role of inter-alpha-inhibitor and its related proteins in urolithiasis. Purification of an inter-alpha-inhibitor related protein from the bovine kidney.
Urine contains several macromolecules that inhibit calcium oxalate (CaOx) crystallization. Among them is bikunin, the light chain of most of the inter-alpha-inhibitor (IalphaI) family of glycoproteins. This study aimed to verify whether bikunin and other members of the IalphaI family are synthesized in the kidneys or derived exclusively from the plasma. Proteins extracted from homogenized bovine kidney were applied successively to three chromatographic steps on DEAE-Sephacel, Sephacryl S-300, and Mono Q column. The inhibitory activity was assayed using a CaOx crystallization system. The presence of IalphaI-related proteins was determined by electrophoresis and Western blotting. The results showed that kidney extract contained a 125-kDa protein that cross-reacted with anti-IalphaI antibodies. This protein inhibited CaOx crystallization efficiently. According to its molecular weight and immunoreaction with anti-IalphaI antibody, the 125-kDa protein could be pre-alpha-inhibitor. The latter is known to encompass a heavy chain and bikunin, which may explain its inhibitory activity against CaOx crystallization. Consequently, we hypothesize that kidneys may produce some IalphaI-related proteins that are involved in the inhibition of stone formation.
Atmani
F
F
Université Mohamed 1er, Faculté des Sciences, Département de Biologie, Oujda-Morocco. atmani@sciences.univ-oujda.ac.ma
Khan
S R
SR
D000510:Alpha-Globulins / Q000302:isolation & purification* / Q000502:physiology*; D000818:Animals; D015153:Blotting, Western; D002129:Calcium Oxalate / Q000037:antagonists & inhibitors; D002417:Cattle; D002845:Chromatography; D003460:Crystallization; D004586:Electrophoresis; D006023:Glycoproteins / Q000302:isolation & purification*; D066298:In Vitro Techniques; D007668:Kidney / Q000737:chemistry*; D008562:Membrane Glycoproteins; D014360:Trypsin Inhibitor, Kunitz Soybean; D014361:Trypsin Inhibitors / Q000302:isolation & purification* / Q000502:physiology*; D014545:Urinary Calculi / Q000517:prevention & control*
D000510:Alpha-Globulins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; C109148:SPINT2 protein, human; D014361:Trypsin Inhibitors; D002129:Calcium Oxalate; C011877:inter-alpha-inhibitor; D014360:Trypsin Inhibitor, Kunitz Soybean