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Purification and Characterization of an Anticalcifying Protein from the Seeds of Trachyspermum ammi (L.)

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Abstract:

Till date various plants extract have been studied to reduce the incidence of urolithiasis but the identification of naturally occurring calcium oxalate (CaOx) inhibitory biomolecules from plants was hampered in past by limitation in identification method. The present study is aimed at examining the efficacy of Trachyspermum ammi on CaOx crystallization in vitro and further by combining conventional biochemical methods with recent advances in mass spectrometry, a novel calcium oxalate (CaOx) crystal growth inhibitor was purified from the seeds of Trachyspermum ammi. An anticalcifying protein from the seeds of Trachyspermum ammi was purified by three step purification scheme; ammonium sulphate fractionation, anion exchange chromatography and molecular sieve chromatography based on its ability to inhibit calcium oxalate crystallization in vitro. An anticalcifying protein having molecular weight 107 kDa and isolectric point 6.2 was isolated. Amino acid analysis of Trachyspermum ammi anticalcifying protein (TAP) showed abundant presence of acidic amino acids (Asp and Glu). Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry of TAP showed similarities with an unnamed protein product of Vitis vinifera (CAO23876) after matching peptide mass fingerprints in MASCOT search engine. Two EF hand domains were identified in unnamed protein product of Vitis vinifera (CAO23876) by SMART normal module. Due to a significant similarity of TAP with unnamed protein product of Vitis vinifera, presence of two EF hand domains in TAP was anticipated, signifying its calcium binding properties which is a feature of most kidney stone inhibitory proteins.





Keywords: Calcium oxalate; Trachyspermum ammi; anticalcifying; inhibitory protein; urolithiasis

Document Type: Research Article

DOI: http://dx.doi.org/10.2174/092986609787316252

Publication date: February 1, 2009

More about this publication?
  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.
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