Identification and characterization of urinary peptides using Q-TOF mass spectrometry in cow
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Identification and characterization of urinary peptides using Q‑TOF mass spectrometry in cow Ambika Sharma1 · Rajesh Nigam1 · Ashish Kumar2 · Abhishek Pal1 Received: 3 April 2020 / Revised: 2 June 2020 / Accepted: 10 June 2020 © Springer Nature Singapore Pte Ltd. 2020
Abstract Cow urine has been found to be the most effective secretion of animal origin with innumerable therapeutic values. It has potent antimicrobial activity which may be attributed to the presence of urinary peptides in it. Lack of evidence for the presence of such antimicrobial peptides (AMPs) in cow urine lead to the present study. Using a technique of membrane filtration, ion exchange chromatography, acid-urea poly-acrylamide gel electrophoresis (AU-PAGE), reverse phase high performance liquid chromatography (RP-HPLC) and ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF-MS/MS), we successfully extracted and characterized cationic peptides from the urine of healthy, normal cycling indigenous cow. We could identify two urinary peptides which were reported to contain antimicrobial properties. These identified peptides were 97 and 99 amino acids long Neutrophil defensin-4 and β-defensin-127 of defensin class. These peptides may be responsible for the innate immunity and serve as an alternative to the conventional antibiotics in the time to come. Keywords Urinary peptides · Neutrophil defensin-4 · β-defensin-127 · Q-TOF–MS/MS · AU-PAGE · Cow
Introduction Antimicrobial peptides (AMPs) are short and generally positively charged (cationic) peptides. AMPs are found in almost all forms of life that includes microorganisms to humans. They have a broad spectrum of activity against microbes ranging from parasites to viruses. Cationic antimicrobial peptides are generally divided into two broad families of defensin and cathelicidin. Defensins are small cysteine rich antimicrobial peptides expressed in various tissues. They are classified into α-, β- and θ-defensins. Defensins have potent antibacterial, antifungal, and cytotoxic activities. In mammals, defensins are molecules of innate and adaptive immunity, with a broad spectrum of antimicrobial activity (Brogden et al. 2003). Defensin molecules possess six highly conserved cysteines (Ganz 2003). α-Defensins are abundant * Ambika Sharma [email protected] 1
Department of Veterinary Biochemistry, College of Veterinary Science and Animal Husbandry, DUVASU, Mathura, Uttar Pradesh 281 001, India
Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), Rishikesh, Uttarakhand 249203, India
2
in neutrophils, paneth cells of the small intestine, whereas β-defensins are generally expressed on epithelial cells lining various organs and also in various biological fluids including urine. θ-Defensins has been identified in rhesus monkey’s leucocytes (Tang 1999). Urine contains proteins and peptides which are either generated in the urinary tract and have specific functions there, or are the filtered or secreted by-pro
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