Proteins in renal stones and urine of stone formers.
Knowledge of the essential characteristics of macromolecules constituting the organic matrix of the nidus of urinary stones is required to understand the mechanism of urolithogenesis. The aim of this study was to isolate and characterise those stone nidus proteins. Using an extraction buffer containing SDS and beta-mercaptoethanol, we were able to overcome known problems of protein isolation from urinary stone matrix. These proteins were characterised by a strong tendency to aggregate under reducing and denaturing conditions. On SDS-PAGE, their molecular weights range from < or = 12 to 66 kDa. Antisera raised against stone matrix proteins showed a cross-reactivity between proteins isolated from different stones irrespective of their origin or mineral composition. Moreover, urinary proteins from stone formers also cross-reacted with these whereas there was no reaction with urinary proteins of non-stone formers. Western blotting confirmed these findings. Given the above summarised properties, it can be safely concluded that these proteins are prevalent in urines of stone formers, that they are selectively incorporated into renal stones of all aetiologies, and that they most likely have a role in nidus and, therefore, early stone formation.
Siddiqui
A A
AA
Department of Biochemistry, The Aga Khan University, Karachi, Pakistan. anwar.siddiqui@aku.edu
Sultana
T
T
Buchholz
N P
NP
Waqar
M A
MA
Talati
J
J
D015153:Blotting, Western; D002021:Buffers; D002129:Calcium Oxalate / Q000032:analysis; D002130:Calcium Phosphates / Q000032:analysis; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007669:Kidney Calculi / Q000737:chemistry*; D008274:Magnesium / Q000032:analysis; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011507:Proteinuria; D014527:Uric Acid / Q000032:analysis; D014556:Urine / Q000737:chemistry
D002021:Buffers; D002130:Calcium Phosphates; D011506:Proteins; C485828:alpha-tricalcium phosphate; C083359:crystal matrix protein, human; C485830:tetracalcium phosphate; D002129:Calcium Oxalate; D014527:Uric Acid; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D008274:Magnesium; C485829:calcium phosphate, dibasic, anhydrous