tailieunhanh - Báo cáo khoa học: Identification of novel fibronectin fragments detected specifically in juvenile urine

Ugl-Y (young age-related urinary glycoprotein) is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. Isoelectric focusing analysis of Ugl-Y gives a set of three bands, Y1, Y2 and Y3, in the pH region around 3. | ỊFEBS Journal Identification of novel fibronectin fragments detected specifically in juvenile urine Reiko Iida1 Toshihiro Yasuda2 and Koichiro Kishi3 1 Division of Forensic Medicine Faculty of MedicalSciences University of Fukui Japan 2 Division of MedicalGenetics and Biochemistry Faculty of MedicalSciences University of Fukui Japan 3 Department of LegalMedicine Gunma University Graduate Schoolof Medicine Japan Keywords age-related extracellular matrix fibronectin fragments MALDI-TOF MS urine Correspondence R. Iida Division of Forensic Medicine Faculty of MedicalSciences University of Fukui Fukui 910-1193 Japan Fax 81 776 61 8108 Tel 81 776 61 8340 E-mail riida@ Received 8 May 2007 revised 28 May 2007 accepted 8 June 2007 doi Ugl-Y young age-related urinary glycoprotein is an age-specific protein that we have previously identified in urine from healthy subjects under 18 years of age. Isoelectric focusing analysis of Ugl-Y gives a set of three bands Y1 Y2 and Y3 in the pH region around 3. To determine the complete structure of Ugl-Y purified Y1 and Y2 from pediatric urine were enzymatically cleaved and the resulting peptides were analyzed by protein sequencing and or MALDI-TOF MS. As a result it was demonstrated that Y1 consists of 189 amino acid residues and is identical to the region from A723 to R911 of fibronectin whereas Y2 consists of 181 amino acid residues and is identical to the region from A723 to R903. Electrophoretic analysis of the lysate prepared from COS-7 cells transfected with Y1- or Y2-expressing vectors gave specific bands corresponding to Y1 or Y2 respectively showing the validity of the sequences determined. Partial purification of pediatric serum followed by western blotting revealed that Ugl-Y is derived from plasma. Furthermore Ugl-Y was generated by in vitro digestion of fibronectin by acid protease in extracts of osteoclast cells. These findings suggest that Ugl-Y is probably produced by .