In veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited (1,2,3). The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an “healthy profile” with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and α2Macroglobulin (170 kDa); at lower molecular weights Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients. Proteomic techniques were successfully used to investigate proteinuria, indicating that two proteins are differentially expressed in urine of healthy and CKD cats. These proteins could be considered as promising biomarkers of chronic renal damage in feline patients. References 1. Lemberger SI, Deeg CA, Hauck SM, Amann B, Hirmer S, Hartmann K, Dorsh R. Comparison of urine protein profiles in cats without urinary tract disease and cats with idiopathic cystitis, bacterial urinary tract infection, or urolithiasis. Am J Vet Res. 2011;72(10):1407-15. 2. Schaefer H, Kohn B, Schweigert FJ, Raila J. Quantitative and qualitative urine protein excretion in dogs with severe inflammatory response syndrome. J Vet Intern Med. 2011;25(6):1292-7. 3. Nabity MB, Lees GE, Dangott LJ, Cianciolo R, Suchodolski JS, Steiner JM. Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease. Vet Clin Pathol. 2011;40(2):222. Figures Figure 1. SDS-PAGE of urine sampples from healthy and CKD cats stained with silver staining. a) Electrophoretic profiles: 1 MW marker; 2-6 CKD urine samples; 7-8, pools of healthy urine samples female and male respectively; b) Pherograms obtained by ImageJ software.
Enea Ferlizza, Aurora Cuoghi, Emanuela Monari, Andrea Neagu, Francesco Dondi, Emilio Carpenè, et al. (2013). Urine proteome from healthy and CKD cats.
Urine proteome from healthy and CKD cats
FERLIZZA, ENEA;DONDI, FRANCESCO;CARPENE', EMILIO;ISANI, GLORIA
2013
Abstract
In veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited (1,2,3). The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an “healthy profile” with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and α2Macroglobulin (170 kDa); at lower molecular weights Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients. Proteomic techniques were successfully used to investigate proteinuria, indicating that two proteins are differentially expressed in urine of healthy and CKD cats. These proteins could be considered as promising biomarkers of chronic renal damage in feline patients. References 1. Lemberger SI, Deeg CA, Hauck SM, Amann B, Hirmer S, Hartmann K, Dorsh R. Comparison of urine protein profiles in cats without urinary tract disease and cats with idiopathic cystitis, bacterial urinary tract infection, or urolithiasis. Am J Vet Res. 2011;72(10):1407-15. 2. Schaefer H, Kohn B, Schweigert FJ, Raila J. Quantitative and qualitative urine protein excretion in dogs with severe inflammatory response syndrome. J Vet Intern Med. 2011;25(6):1292-7. 3. Nabity MB, Lees GE, Dangott LJ, Cianciolo R, Suchodolski JS, Steiner JM. Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease. Vet Clin Pathol. 2011;40(2):222. Figures Figure 1. SDS-PAGE of urine sampples from healthy and CKD cats stained with silver staining. a) Electrophoretic profiles: 1 MW marker; 2-6 CKD urine samples; 7-8, pools of healthy urine samples female and male respectively; b) Pherograms obtained by ImageJ software.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
