Cauxin is an abundant protein in feline urine. We have used proteomics strategies to characterize cauxin from the urine of domestic cats and a number of big cat species. Proteins were resolved by gel-based electrophoretic purification and subjected to in-gel digestion with trypsin. The resultant tryptic peptides were mass-measured by matrix-assisted laser desorption ionization time of flight mass spectrometry. Peptides were also resolved by liquid chromatography and analyzed by electrospray ionization and tandem mass spectrometry to generate fragment ion data to infer the amino acid sequence. We identified cauxin polymorphisms and corrected a sequencing artifact in cauxin from the domestic cat. The proteomics data also provided positive evidence for the presence of a cauxin homolog in the urine of big cats (Pantherinae), including the Sumatran tiger, Asiatic lion, clouded leopard, Persian leopard, and jaguar. The levels of cauxin in the urine of all big cats were substantially lower than that in the urine of intact male domestic cats.
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