Abstract
Demineralization of calcium oxalate (CaOx) crystals precipitated from human urine in vitro yields an organic crystal matrix extract (CME) consisting predominantly of a single protein which we originally named crystal matrix protein but have subsequently shown to be a urinary form of prothrombin activation peptide fragment 1 (F1). The aim of this study was to determine whether CME is a promoter or inhibitor of CaOx crystallization The effect of CME on CaOx crystal growth and aggregation was tested using a standard seeded crystallization system, and its effect quantified by use of particle size analysis and a computer model. In addition, the effect of CME on the crystallization of CaOx was tested in undiluted, ultrafiltered human urine using Coulter Counter analysis and scanning electron microscopy. It was shown that CME is a potent inhibitor of CaOx crystal growth and aggregation in a seeded metastable solution. However, of greater significance is that at a concentration of 10 mg/l it completely reversed the formation of large crystalline aggregates that form upon the removal of urinary macromolecules from undiluted urine. It was concluded that CME is the most potent macromolecular urinary inhibitor yet to be tested in urine in vitro. By preventing the aggregation of newly formed crystals, the components of CME may significantly reduce the probability of particle retention in vivo and therefore the occurrence of urolithiasis.
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References
Boyce WH, Garvey FK (1956) The amount and nature of the organic matrix in urinary calculi: a review. J Urol 76:213
Cabrera N, Vermilyea DA, (1958) In: Doremus RH, Roberts BW, Turnbull D (eds) Growth perfection of crystals. Wiley, New York, p 393
Doyle IR. The role of urinary proteins in calcium oxalate urolithiasis. PhD thesis, The Flinders University of South Australia 1992
Doyle IR, Ryall RL, Marshall VR (1989) The effect of low-speed centrifugation and millipore filtration on the urinary protein content. In: Walker VR, Sutton RAL, Cameron ECB, Pak CYC, Robertson WG (eds) Urolithiasis. Plenum Press, New York, p 593
Doyle IR, Ryall RL, Marshall VR (1991) The incorporation of urinary protein into calcium oxalate crystals precipitated from human urine: A highly selective phenomenon. Clin Chem 37:1589
Drach GW, Thorson S, Randolph A (1980) Effects of urinary organic macromolecules on crystallization of calcium oxalate: enhancement of nucleation. J Urol 123:519
Edyvane KA, Hibberd CM, Harnett RM, Marshall VR, Ryall RL (1987) Macromolecules inhibit calcium oxalate crystal growth in whole human urine. Clin Chim Acta 167:329
Deyvane KA, Ryall RL, Marshall VR (1986) The influence of serum and serum proteins on calcium oxalate crystal growth and aggregation. Clin Chim Acta 157:81
Edyvane KA, Ryall RL, Mazzachi RD, Marshall VR (1987) The effect of serum on the crystallization of calcium oxalate in whole human urine: inhibition disguised as apparent promotion. Urol Res 15:87
Fleisch H (1978). Inhibitors and promoters of stone formation. Kidney Int 13:361
Grover PK, Ryall RL, Marshall VR (1990) Does Tamm-Horsfall mucoprotein inhibit or promote calcium oxalate crystallization in human urine? Clin Chim Acta 190:223
Hallson PC, Rose GA (1979) Uromucoids and urinary stone formation. Lancet 1:1000
Hess B, Nakagawa Y, Coe FL (1989) Nephrocalcin isolated from human kidney stones is a defective calcium-oxalate-monohydrate crystal-aggregation inhibitor. In: Walker VR, Sutton RAL, Cameron ECB, Pak CYC, Robertson WG (eds) Urolithiasis. Plenum Press, New York, p 137
Hoyer JR (1994) Role of uropontin in calcium stone formation. In: Ryall RL, Bais R, Marshall VR, Rofe AM, Smith LH, Walker VR (eds) Urolithiasis II. Plenum Press, New York, p 253
Iwata H, Kamei O, Abe Y, Nishio S, Wakatsuki A, Ochi K, Takeuchi M (1988) The organic matrix of urinary uric acid crystals. J Urol 139:607
Koide T, Takemoto M, Itatani H, Takaha M, Sonoda T (1981) Urinary macromolecular substances as natural inhibitors of calcium oxalate crystal aggregation. Invest Urol 18:382
Koide T, Yoshioka T, Yamaguchi S, Hosokawa S, Utsunomiya M, Sonoda T (1990) Urinary crystal surface binding substances on calcium oxalate crystals. Urol Res 18:387
Lian JB, Prien EL Jr, Glimcher MJ, Gallop PM (1977) The presence of protein-bound γ-carboxyglutamic acid in calcium-containing renal calculi. J Clin Invest 59:1151
Morse RM, Resnick MI (1988) A new approach to the study of urinary macromolecules as participant in calcium oxalate crystallization. J Urol 139:869
Morse RM, Resnick MI (1989) A study of the incorporation of urinary macromolecules into crystals of different mineral composition. J Urol 141:641
Nakagawa Y, Abram F, Kézdy J, Kaiser ET, Coe FL (1983) Purification and characterization of the principal inhibitor of calcium oxalate monohydrate crystal growth in human urines. J Biol Chem 258:12594
Nakagawa Y, Netzer M, Coe FL (1990) Immunohistochemical localization of nephrocalcin (NC) to proximal tubule and thick ascending limb of Henle's loop (TAHL) of human and mouse kidney: resolution of a conflict. Kidney Int 37:A474
Nishio S, Wakatsuki A, Iwata H, Matsumoto A, Takeuchi M (1989) Urinary macromolecular-inhibitory-substances of calcium-oxalate-monohydrate crystal growth. In: Walker VR, Sutton RAL, Cameron ECB, Pak CYC, Robertson WG (eds) Urolithiasis. Plenum Press, New York, p 235
Petersen TE, Thøgerson I, Petersen SE (1989) Identification of hemoglobin and two serine proteases in acid extracts of calcium containing kidney stones. J Urol 142:176
Resnick MI, Gammon CW, Sorrell MB, Boyce WH (1980) Calcium-binding proteins and renal lithiasis. Surgery 88:239
Resnick MI, Sorrell ME, Bailey JA, Boyce WH (1982) Inhibitory effects of urinary calcium-binding substances and calcium oxalate crystallization J Urol 127:568
Robertson WG, Peacock M (1972) Calcium oxalate crystalluria and inhibitors of crystallisation in recurrent renal stone-formers. Clin Sci 43:499
Robertson WG, Scurr DS (1986) Modifiers of calcium oxalate crystallization found in urine. I. Studies with a continuous crystallizer using an artificial urine. J Urol 135:1322
Robertson WG, Peacock M, Nordin BEC (1968) Activity products in stone-forming and non-stone-forming urine. Clin Sci 34:579
Romberg RW, Werness PG, Riggs BL, Mann KG (1986) Inhibition of hydroxyapatite crystal growth by bone-specific and other calcium-binding proteins. Biochemistry 25:1176
Ryall RL, Bagley C, Marshall VR (1981) Independent assessment of the growth and aggregation of calcium oxalate crystals using the Coulter Counter. Invest Urol 18:401
Ryall RL, Harnett RM, Marshall VR (1981) The effect of urine, pyrophosphate, citrate, magnesium and glycosaminoglycans on the growth and aggregation of calcium oxalate crystals in vitro. Clin Chim Acta 112:349
Ryall RL, Hibberd CM, Marshall VR (1985) A method for studying inhibitiory activity in whole urine. Urol Res 13:285
Ryall RG, Ryall RL, Marshall VR (1986) A computer model for the determination of extents of growth and aggregation of crystals from changes in their size distribution. J Cryst Growth 76:290
Ryall RL, Harnett RM, Hibberd CM, Edyvane KA, Marshall VR (1991) Effects of chondroitin sulphate, human serum albumin and Tamm-Horsfall mucoprotein on calcium oxalate crystallization in undiluted urine. Urol Res 19:181
Ryall RL, Ryall RG, Doyle IR, Marshall VR (1993) The dependence of crystal growth on the state of crystal aggregation: Implications for urolithiasis research. J Crystal Growth 33:87
Scurr DS, Robertson WG (1986) Modifiers of calcium oxalate crystallization in urine. II. Studies on their mode of action in an artificial urine. J Urol 136:128
Shiraga H, Min W, VanDusen WJ Clayman MD, Miner D, Terrell CH, Sherbotie JR, Foreman JW, Przysiecki C, Neilson EG, Hoyer JR (1992) Inhibition of calcium oxalate crystal growth in vitro by uropontin: Another member of the aspartic acid-rich protein superfamily. Proc Natl Acad Sci USA 89:426
Sørenson S, Hansen K, Bak S, Justesen SJ (1990) An unidentified macromolecular inhibitory constituent of calcium oxalate crystal growth in human urine. Urol Res 18:373
Stapleton AMF, Ryall (in press) Blood coagulation proteins and urolithiasis are linked: Crystal matrix protein is the F1 activation peptide of human prothrombin Br J Urol
Stapleton AMF, Seymour AE, Brennan JS, Doyle IR, Marshall VR, Ryall RL, (1993) The immunohistochemical distribution and quantification of crystal matrix protein. Kidney Int 44:817
Stapleton AMF, Simpson RJ, Ryall RL (1993) Crystal matrix protein is related to human prothrombin. Biochem Biophys Res Commun 195:1199
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Doyle, I.R., Marshall, V.R., Dawson, C.J. et al. Calcium oxalate crystal matrix extract: the most potent macromolecular inhibitor of crystal growth and aggregation yet tested in undiluted human urine in vitro. Urol. Res. 23, 53–62 (1995). https://doi.org/10.1007/BF00298852
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DOI: https://doi.org/10.1007/BF00298852