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. 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, revealing different protein patterns between healthy and CKD cats. Some of these proteins could be considered as promising biomarkers of chronic renal damage in feline patients.
Proteomic analysis of urine from healthy and CKD cats / Enea Ferlizza; Aurora Cuoghi; Emanuela Monari; Andrea Neagu; Francesco Dondi; Emilio Carpenè; Gloria Isani. - STAMPA. - (2013), pp. 101-101. (Intervento presentato al convegno ItPA, 8th Annual National Conference tenutosi a Padova nel 18-21 giugno 2013).
Proteomic analysis of urine 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. 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, revealing different protein patterns between healthy and CKD cats. Some of these proteins could be considered as promising biomarkers of chronic renal damage in feline patients.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
