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Peeping into human renal calcium oxalate stone matrix: characterization of novel proteins involved in the intricate mechanism of urolithiasis.

Aggarwal KP, Tandon S, Naik PK, Singh SK, Tandon C - PLoS ONE (2013)

Bottom Line: Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal-membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure.Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level.Thus, these proteins having potential to modulate calcium oxalate crystallization will throw light on understanding and controlling urolithiasis in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh, India.

ABSTRACT

Background: The increasing number of patients suffering from urolithiasis represents one of the major challenges which nephrologists face worldwide today. For enhancing therapeutic outcomes of this disease, the pathogenic basis for the formation of renal stones is the need of hour. Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal-membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure.

Methods: Proteins were isolated from the matrix of human CaOx containing kidney stones. Proteins having MW>3 kDa were subjected to anion exchange chromatography followed by molecular-sieve chromatography. The effect of these purified proteins was tested against CaOx nucleation and growth and on oxalate injured Madin-Darby Canine Kidney (MDCK) renal epithelial cells for their activity. Proteins were identified by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF MS) followed by database search with MASCOT server. In silico molecular interaction studies with CaOx crystals were also investigated.

Results: Five proteins were identified from the matrix of calcium oxalate kidney stones by MALDI-TOF MS followed by database search with MASCOT server with the competence to control the stone formation process. Out of which two proteins were promoters, two were inhibitors and one protein had a dual activity of both inhibition and promotion towards CaOx nucleation and growth. Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level.

Conclusions: We identified and characterized Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, RIMS-binding protein 3A, Macrophage-capping protein as novel proteins from the matrix of human calcium oxalate stone which play a critical role in kidney stone formation. Thus, these proteins having potential to modulate calcium oxalate crystallization will throw light on understanding and controlling urolithiasis in humans.

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Related in: MedlinePlus

HPLC analysis.HPLC analysis of (D) NP2; (E) OP1.
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pone-0069916-g009: HPLC analysis.HPLC analysis of (D) NP2; (E) OP1.

Mentions: The protein bands detected in fractions MP1, MP2, NP1, NP2 and OP1 were excised, in-gel trypsin digested and identified by matrix assisted laser desorption/ionization–time of flight (MALDI-TOF) MS. Purity of the samples was tested using RP-HPLC (Figure 8, 9). Using the Mascot search engine (http://www.matrixscience.com), the MALDI-TOF data obtained from fractions MP1, MP2, NP1, NP2, OP1 matched significantly with Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, Macrophage-capping protein and RIMS-binding protein 3A respectively (Table 1). MASCOT search engine revealed that these proteins had MOWSE score of 26, 31, 23, 26 and 23 respectively and with good sequence coverage.


Peeping into human renal calcium oxalate stone matrix: characterization of novel proteins involved in the intricate mechanism of urolithiasis.

Aggarwal KP, Tandon S, Naik PK, Singh SK, Tandon C - PLoS ONE (2013)

HPLC analysis.HPLC analysis of (D) NP2; (E) OP1.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3722206&req=5

pone-0069916-g009: HPLC analysis.HPLC analysis of (D) NP2; (E) OP1.
Mentions: The protein bands detected in fractions MP1, MP2, NP1, NP2 and OP1 were excised, in-gel trypsin digested and identified by matrix assisted laser desorption/ionization–time of flight (MALDI-TOF) MS. Purity of the samples was tested using RP-HPLC (Figure 8, 9). Using the Mascot search engine (http://www.matrixscience.com), the MALDI-TOF data obtained from fractions MP1, MP2, NP1, NP2, OP1 matched significantly with Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, Macrophage-capping protein and RIMS-binding protein 3A respectively (Table 1). MASCOT search engine revealed that these proteins had MOWSE score of 26, 31, 23, 26 and 23 respectively and with good sequence coverage.

Bottom Line: Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal-membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure.Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level.Thus, these proteins having potential to modulate calcium oxalate crystallization will throw light on understanding and controlling urolithiasis in humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh, India.

ABSTRACT

Background: The increasing number of patients suffering from urolithiasis represents one of the major challenges which nephrologists face worldwide today. For enhancing therapeutic outcomes of this disease, the pathogenic basis for the formation of renal stones is the need of hour. Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal-membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure.

Methods: Proteins were isolated from the matrix of human CaOx containing kidney stones. Proteins having MW>3 kDa were subjected to anion exchange chromatography followed by molecular-sieve chromatography. The effect of these purified proteins was tested against CaOx nucleation and growth and on oxalate injured Madin-Darby Canine Kidney (MDCK) renal epithelial cells for their activity. Proteins were identified by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF MS) followed by database search with MASCOT server. In silico molecular interaction studies with CaOx crystals were also investigated.

Results: Five proteins were identified from the matrix of calcium oxalate kidney stones by MALDI-TOF MS followed by database search with MASCOT server with the competence to control the stone formation process. Out of which two proteins were promoters, two were inhibitors and one protein had a dual activity of both inhibition and promotion towards CaOx nucleation and growth. Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level.

Conclusions: We identified and characterized Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, RIMS-binding protein 3A, Macrophage-capping protein as novel proteins from the matrix of human calcium oxalate stone which play a critical role in kidney stone formation. Thus, these proteins having potential to modulate calcium oxalate crystallization will throw light on understanding and controlling urolithiasis in humans.

Show MeSH
Related in: MedlinePlus