Anti-dsDNA autoantibody cross-reacts with the C-terminal hydrophobic cluster region containing phenylalanines in the acidic ribosomal phosphoprotein P1 to exert a cytostatic effect on the cells.
The present study was an attempt to map the epitope in P1 protein which may cross-react with anti-dsDNA. In addition to wild-type P1, the genes of its C-terminal mutants were cloned and expressed. The binding activity of these proteins with anti-dsDNA was determined by Western blot and ELISA. The P1 mutants with complete deletion of the acidic charge and hydrophobic clusters, deletion of the hydrophobic cluster, or replacement of the phenylanlanines with alanine in the hydrophobic cluster lost the binding activity. Moreover, P1 mutants with mutation at the serine phosphorylation sites (positions 102 and 105) retained their binding activities with anti-dsDNA. However, those with mutation at the serine phosphorylation sites and without the hydrophobic cluster lost their binding activities. These findings suggest that phenylalanines in the C-terminal hydrophobic cluster region of ribosomal P proteins are essential to their cross-reactivity with anti-dsDNA.
Sun
K H
KH
Faculty of Medical Technology & Institute of Biotechnology in Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China. khsun@dns.ym.edu.tw
Hong
C C
CC
Tang
S J
SJ
Sun
G H
GH
Liu
W T
WT
Han
S H
SH
Yu
C L
CL
D000595:Amino Acid Sequence; D000818:Animals; D001323:Autoantibodies / Q000276:immunology* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA / Q000276:immunology*; D018604:Epitope Mapping; D005920:Glomerular Mesangium / Q000166:cytology / Q000276:immunology*; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008969:Molecular Sequence Data; D010649:Phenylalanine / Q000276:immunology*; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D051381:Rats; D012269:Ribosomal Proteins / Q000235:genetics / Q000276:immunology*
D001323:Autoantibodies; D010750:Phosphoproteins; D012269:Ribosomal Proteins; C035690:ribosomal phosphoprotein P1; D010649:Phenylalanine; D004247:DNA