Isolation and partial characterization of crystal matrix protein as a potent inhibitor of calcium oxalate crystal aggregation: evidence of activation peptide of human prothrombin

Suzuki, Kôji; Moriyama, Manabu; Nakajima, Chiaki; Kawamura, K; Miyazawa, Katsuhito; Tsugawa, Ryuzo; Kikuchi, Norihisa; Nagata, Kiyoshi

doi:10.1007/bf00431548

Cited by 43 publications

(37 citation statements)

References 17 publications

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“…The inhibitory effects of osteopontin and prothrombin fragment 1 on the crystallization and aggregation of CaOx crystals have been reported. 25,26 Protein Z, protein S, and prothrombin, which are vitamin K-dependent calcium-binding proteins, are thought to act as regulators in the coagulation process. 27,28 These proteins have a Gla-containing domain that is able to chelate calcium ions.…”

Section: Proteins Identified In Stones Using Lc-ms/msmentioning

confidence: 99%

Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate

et al. 2015

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In this study, we performed proteomic analysis following sequential protein extraction on calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) urinary stones to determine the specific matrix proteins according to the crystal components of the stones. After X-ray and IR analysis of 13 urinary stones, matrix proteins were sequentially extracted with KCl, formic acid, guanidine-HCl, and EDTA, before SDS-electrophoresis followed by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The electrophoretic patterns of the extracted proteins differed from that of COM and COD stones. LC-MS/MS identified 65 proteins, of which many were cellular plasma proteins, and were frequently detected regardless of the crystal components. However, 6 proteins (protein Z, protein S, prothrombin, osteopontin, fatty acid binding protein 5, and ubiquitin) were detected in the final EDTA fractions of COM stones. These proteins are involved in the coagulation process or osteometabolism, and thus the roles they play are of particular interest.

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Section: Proteins Identified In Stones Using Lc-ms/msmentioning

confidence: 99%

Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate

et al. 2015

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“…Blots were stained with Coomassie blue (0.1%), destained with 50% methanol and 10% acetic acid, and the predominant band at 39 kDa was excised. Amino acid compositional analysis, performed at the University of Chicago Amino Acid and Protein Core Lab, is shown in Table 1, in which CAI is compared with three other urinary crystallization inhibitors, uropontin (UP) (31), human urinary prothrombin fragment 1 (UPTF1) (33), and bikunin (34). Whereas the estimated total net charges on the protein core of UP (Ϫ25.5), UPTF1 (Ϫ14.3), and bikunin (Ϫ7.4) are each negative, CAI (Ϫ2.1) was nearly neutral.…”

Section: Cultured Renal Cells Constitutively Release Cai Binding Of mentioning

confidence: 99%

Renal epithelial cells constitutively produce a protein that blocks adhesion of crystals to their surface

Kumar

Farell

et al. 2004

American Journal of Physiology-Renal Physiology

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. Renal epithelial cells constitutively produce a protein that blocks adhesion of crystals to their surface. Am J Physiol Renal Physiol 287: F373-F383, 2004. First published April 20, 2004 10.1152/ajprenal.00418.2003.-Attachment of newly formed crystals to renal tubular epithelial cells appears to be a critical step in the development of kidney stones. The present study was undertaken to identify autocrine factors released from renal epithelial cells into the culture medium that inhibit adhesion of calcium oxalate crystals to the cell surface. A 39-kDa glycoprotein that is constitutively secreted by renal cells was purified by gel filtration chromatography. Amino acid microsequencing revealed that it is novel and not structurally related to known inhibitors of calcium oxalate crystallization. Hence, it was named crystal adhesion inhibitor, or CAI. Immunoreactive CAI was detected in diverse rat tissues, including kidney, heart, pancreas, liver, and testis. Immunohistochemistry revealed that CAI is present in the renal cell cytosol and is also on the plasma membrane. Importantly, CAI is present in normal human urine, from which it can be purified using calcium oxalate monohydrate crystal affinity chromatography. CAI could be an important defense against crystal attachment to tubular cells and the subsequent development of renal stones in vivo.calcium oxalate monohydrate; cell-crystal interaction; DING protein; inhibitor; nephrolithiasis NEPHROLITHIASIS IS AN EXTREMELY common condition in the United States, affecting up to 10% of the population at some point during their lives (26,32). Although many affected individuals have identifiable urinary metabolic risk factors, such as excessively concentrated urine that may contain too much calcium, uric acid, or oxalate, or perhaps too little citrate, many do not (12). Therefore, the concentration of these urinary constituents does not appear to fully explain the formation of renal stones. In addition, nucleation and growth of individual crystals appear unlikely to produce particles large enough to occlude the nephron lumen in vivo (15). Recent evidence suggests, that in many calcium oxalate stone formers, the earliest changes may be depositions of calcium phosphate in the medullary interstitium, which then serve as a nidus for a calcium oxalate stone (14). The processes that mediate calcium phosphate deposition and its evolution into calcium oxalate stones remain to be determined. In more marked hyperoxaluric states (e.g., enteric or primary hyperoxaluria), direct adhesion of calcium oxalate crystals to renal epithelial cells may predominate (14). Therefore, our laboratory (19 -24) and others (29,30,37,38,43) have sought to identify regulatory mechanism(s) by which urinary calcium oxalate and calcium phosphate crystals in tubular fluid bind to renal epithelial cells, are retained in the kidney, and become a nidus for stone formation.Previous studies have demonstrated that anionic molecules in tubular fluid can coat calcium oxalate monohydrate (COM) and hydroxyapatite (H...

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“…First of all, it is worthwhile to mention that crystals were washed only with distilled water rather than sodium hydroxide solution as described by some investigators [10][11][12][13]. Recently, we have reported that this procedure of washing contributes to the loss of most THP and albumin [14].…”

Section: Analysis Of Crystal Matrix Proteinsmentioning

confidence: 99%

“…In this regard, it was proposed to analyze proteins extracted from calcium oxalate (CaOx) crystals induced in vitro in normal urine [10][11][12]. The results showed a highly selective incorporation of prothrombin fragment 1 [12,13]. Subsequently, it was concluded that this protein plays an important role in urinary stone formation.…”

Section: Introductionmentioning

confidence: 99%

Quantification of Proteins Extracted from Calcium Oxalate and Calcium Phosphate Crystals Induced in vitro in the Urine of Healthy Controls and Stone-Forming Patients

Atmani

Khan

2002

Urol Int

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We have previously identified proteins extracted from calcium oxalate (CaOx) and calcium phosphate (CaP) crystals generated experimentally in vitro in whole urine of healthy controls and stone formers. No significant differences were detected between protein components in matrices of crystals obtained from both groups. The aim of the present study was to estimate the amounts of six proteins identified earlier in order to investigate the differences, if any, between healthy controls and lithiasis patients. CaOx and CaP crystals were generated in the urine samples by adding an oxalate and phosphate load, respectively. Crystals were harvested, washed, dried, and analyzed. Crystal matrix protein was extracted by demineralizing crystals with EDTA solution, analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and identified immunochemically using Western blot analysis. The quantity of each protein was estimated by densitometric analysis. The predominant proteins found in organic matrices of CaOx crystals induced in the urine of healthy controls were prothrombin-related proteins followed by albumin and osteopontin. In matrices of CaP crystals, the principal proteins were Tamm-Horsfall protein followed by albumin, prothrombin-related proteins and osteopontin. However, when crystals were induced in the urine of stone formers, albumin was the major component of the organic matrix of both CaOx and CaP crystals. We concluded that a subset of urinary proteins is present in crystal matrix. Among them, albumin seems to play a crucial role in kidney stone formation.

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Isolation and partial characterization of crystal matrix protein as a potent inhibitor of calcium oxalate crystal aggregation: evidence of activation peptide of human prothrombin

Cited by 43 publications

References 17 publications

Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate

Proteomic Analysis after Sequential Extraction of Matrix Proteins in Urinary Stones Composed of Calcium Oxalate Monohydrate and Calcium Oxalate Dihydrate

Renal epithelial cells constitutively produce a protein that blocks adhesion of crystals to their surface

Quantification of Proteins Extracted from Calcium Oxalate and Calcium Phosphate Crystals Induced in vitro in the Urine of Healthy Controls and Stone-Forming Patients

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