~ 2393 ~ International Journal of Chemical Studies 2019; 7(1): 2393-2397 P-ISSN: 2349–8528 E-ISSN: 2321–4902 IJCS 2019; 7(1): 2393-2397 © 2019 IJCS Received: 11-11-2018 Accepted: 15-12-2018 Maninder Singh Sheoran Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India Priyanka Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India Ramkaran Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India Ashok Balhara Department of Animal Physiology and Reproduction, CIRB, Hisar, Haryana, India Varij Nayan Department of Animal Physiology and Reproduction, CIRB, Hisar, Haryana, India Sandeep Kumar Senior Scientist, Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India Correspondence Sandeep Kumar Senior Scientist, Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India Biochemical characterization of Urine of Hariana breed of Cattle with special emphasis on presence of various proteins/peptides Maninder Singh Sheoran, Priyanka, Ramkaran, Ashok Balhara, Varij Nayan and Sandeep Kumar Abstract Urine is a non-invasive source of biological fluid, which reflects the physiological status of the animals. We have profiled the cow urinary proteome and analyzed its functional significance. The urine collected from two healthy Hariana cows was concentrated by diafiltration (DF) followed by protein extraction using TCA acetone precipitation. The quality of the protein was assessed by two-dimensional gel electrophoresis (2DE) followed by In-gel digestion method. Functional significance of few selected proteins seems to play important role in normal body functions. This study reports the presence of proteins in urine of healthy Hariana cattle for the first time. Keywords: urine, peptide, cow, two-dimensional gel electrophoresis Introduction Urine contains abundant biomolecules including proteins, peptides and metabolites which are the byproducts of the physiological events occuring in the organism. Urine is formed via glomerular filtration of plasma in the kidneys, which act as a filter to retain most of the plasma proteins (Thongboonkerd et al, 2002) [21] . Though, many low molecular weight proteins and peptides pass through the glomerular membrane, are catabolized in the proximal tubule and then secreted in the urine. In addition to this, abundant serum proteins like albumin, immunoglobulin light chain, transferrin, myoglobin, and receptor-associated protein, after passing through the glomeruli, are reabsorbed by endocytic receptors in the proximal renal tubules (Kerr et al, 1998, Clark et al, 1998) [11, 4] . Overall, the protein concentration in normal urine is usually very low (Adachi et al, 2006) [1] . Moreover, there is interest in using urine for diagnostic applications due to its simple and non-invasive collection compared to other biological fluids. In particular, many systemic diseases may cause a change in the composition of the urinary constituents, which might be useful for diagnosis and disease monitoring (Adachi et al, 2006) [1] . In humans, biomarkers in urine have been used for the diagnosis of kidney and urinary tract diseases (Noiri et al, 2009) [14] . However, very few studies have been reported on the bovine urinary proteome as a potential source for biomarker discovery in disease and pregnancy (Shaked et al, 2001, Simon et al, 2008) [17] . The human urinary proteome has been extensively studied by a variety of methods including two-dimensional gel electrophoresis (2DE) followed by its characterization using matrix assisted laser desorption ionization (MALDI) and liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) (Pieper et al, 2004) [15] . In one study, (Sun et al, 2005) [20] identified 226 unique human proteins in urine using one-dimensional gel electrophoresis (1D- GE) and multidimensional LC-MS/MS. However, to date, no information is available on the bovine urinary proteome. The aim of the present study was to profile the urinary proteome of Hariana cows. The Hariana breed has adapted well to tropical climate and plays important role in milk production in India (Gurnani et al, 1986) [7] . In this proteomic study, a combination of extraction procedures was used to isolate urinary proteins from Hariana cows which were then characterized by MS leading to the identification of a many useful proteins. This is the first report of urinary proteome of Hariana cow and will provide a reference database for future studies in biomarker discovery linked to diseases and associated physiological events.