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A Novel Calcium Oxalate Crystal Growth Inhibitory Protein from the Seeds of Dolichos biflorus (L.)

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Abstract

Recurrence and persistent side effects of present day treatment for urolithiasis restrict their use, so an alternate, using phytotherapy is being sought. Dolichos biflorus seeds, which are used as dietary food in India, possess antilithiatic properties. In the present study, a novel dimeric antilithiatic protein (98 kDa) from its seeds was purified based on its ability to inhibit calcium oxalate crystallization in vitro. Amino acid analysis of Dolichos biflorus antilithiatic protein showed abundant acidic amino acids. The mascot search engine presented sequence similarity with a calcium binding protein, calnexin of Pisum sativum from the m/z data obtained by MALDI TOF mass spectrometer. Above results demonstrate the anticalcifying/antilithiatic nature of a novel protein from the seeds of Dolichos biflorus and thus open new vistas for using plant proteins as therapeutic agents to treat urolithiasis.

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Abbreviations

CNX:

Calnexin

DAP:

Dolichos biflorus antilithiatic protein

CaOx:

Calcium oxalate

CO:

Calcium oxalate monohydrate

C:

Calcium oxalate dihydrate

FT:

Fourier transform infrared spectroscopy

MALDI TOF:

Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry

RP:

Reverse phase

HPLC:

High pressure liquid chromatography

TFA:

Trifluoroacetic acid

EG:

Ethylene glycol

PMSF:

Phenylmethane sulphonyl fluoride

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Acknowledgments

The authors would like to thank Dr. S.·K. Singh, Department of Urology, Post Graduate Institute of Medical and Education research (PGIMER), Chandigarh, India for providing clinical calcium oxalate stones; Dr. Girish Sahni, Director, Institute of Microbial Technology (IMTECH), Chandigarh, India for amino acid analysis; Dr. P. Rama Rao, Director, National Institute of Pharmaceutical Education and research (NIPER), SAS Nagar, India for MALDI-TOF MS.

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Authors and Affiliations

  1. Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173215, India

    Rakesh K. Bijarnia, Tanzeer Kaur & Chanderdeep Tandon

  2. Department of Biochemistry, Panjab University, Chandigarh, India

    Surinder K. Singla

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  1. Rakesh K. Bijarnia
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  2. Tanzeer Kaur
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  3. Surinder K. Singla
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  4. Chanderdeep Tandon
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Correspondence to Chanderdeep Tandon.

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Bijarnia, R.K., Kaur, T., Singla, S.K. et al. A Novel Calcium Oxalate Crystal Growth Inhibitory Protein from the Seeds of Dolichos biflorus (L.). Protein J 28, 161–168 (2009). https://doi.org/10.1007/s10930-009-9179-y

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