Monoclonal antibody RYSK173 recognizes the dinuclear Zn center of serum carnosinase 1 (CN-1)

• Verfasst von: Zhang, Shiqi [VerfasserIn]
• Verfasst von: Lindner, Holger A. [VerfasserIn]
• Verfasst von: Krämer, Bernhard [VerfasserIn]
• Verfasst von: Yard, Benito A. [VerfasserIn]
• Verfasst von: Hauske, Sibylle J. [VerfasserIn]
• Titel: Monoclonal antibody RYSK173 recognizes the dinuclear Zn center of serum carnosinase 1 (CN-1)
• Titelzusatz: possible consequences of Zn binding for CN-1 recognition by RYSK173
• Verf.angabe: Shiqi Zhang, Holger A. Lindner, Sarah Kabtni, Jaap van den Born, Stephan Bakker, Gerjan Navis, Bernard Krämer, Benito Yard, Sibylle Hauske
• E-Jahr: 2016
• Jahr: January 22, 2016
• Umfang: 12 S.
• Fussnoten: Gesehen am 23.01.2019
• Titel Quelle: Enthalten in: PLOS ONE
• Ort Quelle: San Francisco, California, US : PLOS, 2006
• Jahr Quelle: 2016
• Band/Heft Quelle: 11(2016), 1, Artikel-ID e0146831, Seite 1-12
• ISSN Quelle: 1932-6203
• DOI: doi:10.1371/journal.pone.0146831
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Chronic kidney disease
• Sach-SW: Enzyme-linked immunoassays
• Sach-SW: Epitope mapping
• Sach-SW: Medical dialysis
• Sach-SW: Monoclonal antibodies
• Sach-SW: Recombinant proteins
• Sach-SW: Synthetic peptides
• Sach-SW: Zinc
• K10plus-PPN: 1586509640

Abstract

Background and Aims The proportion of serum carnosinase (CN-1) recognized by RYSK173 monoclonal antibody negatively correlates with CN-1 activity. We thus hypothesized that the epitope recognized by RYSK173 is accessible only in a catalytically incompetent conformation of the zinc dependent enzyme and we mapped its position in the CN-1 structure. Since patients with kidney failure are often deficient in zinc and other trace elements we also assessed the RYSK173 CN-1 proportion in serum of these patients and studied the influence of hemodialysis hereon in relation to Zn2+ and Cu2+ concentration during hemodialysis. Methods and Results Epitope mapping using myc-tagged CN-1 fragments and overlapping peptides revealed that the RYSK173 epitope directly contributes to the formation of the dinuclear Zn center in the catalytic domain of homodimeric CN-1. Binding of RYSK173 to CN-1 was however not influenced by addition of Zn2+ or Cu2+ to serum. In serum of healthy controls the proportion of CN-1 recognized by RYSK173 was significantly lower compared to end-stage renal disease (ESRD) patients (1.12 ± 0.17 vs. 1.56 ± 0.40% of total CN-1; p<0.001). During hemodialysis the relative proportion of RYSK173 CN-1 decreased in parallel with increased serum Zn2+ and Cu2+ concentrations after dialysis. Conclusions Our study clearly indicates that RYSK173 recognizes a sequence within the transition metal binding site of CN-1, thus supporting our hypothesis that metal binding to CN-1 masks the epitope. The CN-1 RYSK173 proportion appears overall increased in ESRD patients, yet it decreases during hemodialysis possibly as a consequence of a relative increase in transition metal bound enzyme.