Zinc ion dependent B-cell epitope, associated with primary Sjogren's syndrome, resides within the putative zinc finger domain of Ro60kD autoantigen: physical and immunologic properties.
The Ro/La ribonucleoprotein (RNP) complex is composed of the proteins Ro60kD, Ro52kD, and La48kD that are in association with one small cytoplasmic RNA (YRNA). Specific protein-RNA and protein-protein interactions are thought to occur through the RNP and zinc-finger secondary structure elements of the Ro60kD protein. The aim of our study was to investigate the antigenic properties of the zinc finger domain of the Ro60KD autoantigen and its contribution to the formation of Ro/La RNP complex. It was found that the peptide VSLVCEKLCNEKLLKKARIHPFHILIA (Zif-1), which corresponds to the natural sequence of the zinc finger domain (301-327), and the peptide C(Acm)NEKLLKKARIC(Acm), analogous to the intermediate loop 310-319 (Zif-3) of the same domain of Ro60KD, are recognized by the majority of anti-Ro/SSA and anti-La/SSB positive sera (82.6% and 77.1%, respectively) in the absence of zinc ions. The same sera failed to react with Zif-1 peptide in the presence of Zn2+. In contrast, the addition of zinc ions was necessary for the binding of Zif-1 to recombinant Ro52KD as shown by direct binding experiments of the recombinant protein with synthetic peptides. Our data suggest the zinc finger domain of Ro60kD contains a B-cell epitope with high specificity for primary Sjogren's syndrome. Furthermore, depending on the presence of zinc ions, the zinc finger domain of the Ro60KD protein can exist in two different conformational states favoring either an interaction with the Ro52KD protein or binding with autoantibodies.
Routsias
John G
JG
Department of Pathophysiology, School of Medicine, University of Athens, 11527 Athens, Greece.
Kosmopoulou
Aggeliki
A
Makri
Athina
A
Panou-Pomonis
Eugenia
E
Sakarellos
Constantinos
C
Sakarellos-Daitsiotis
Maria
M
Moutsopoulos
Haralampos M
HM
Tzioufas
Athanasios G
AG
D001323:Autoantibodies / Q000097:blood / Q000737:chemistry / Q000276:immunology; D001324:Autoantigens / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002413:Cations, Divalent; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D011485:Protein Binding; D011487:Protein Conformation; D020733:RNA, Small Cytoplasmic / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry; D012261:Ribonucleoproteins / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D012859:Sjogren's Syndrome / Q000276:immunology*; D021241:Spectrometry, Mass, Electrospray Ionization; D015032:Zinc / Q000737:chemistry / Q000276:immunology*; D016335:Zinc Fingers; D000099207:SS-A Antigen
D001323:Autoantibodies; D001324:Autoantigens; D002413:Cations, Divalent; D000939:Epitopes; D010455:Peptides; D020733:RNA, Small Cytoplasmic; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D015032:Zinc; C116006:RO60 protein, human; D000099207:SS-A Antigen