As an acute-phase protein, human C-reactive protein (CRP) is clinically important. CRPs were purified from several samples in six different pathological conditions, where their levels ranged from 22 to 342 μg/ml. Small, but significant, variations in electrophoretic mobilities on native PAGE suggested differences in molecular mass, charge and/or shape. Following separation by SDS/PAGE, they showed single subunits with some differences in their molecular masses ranging between 27 and 30.5 kDa, but for a particular disease, the mobility was the same for CRPs purified from multiple individuals or pooled sera. Isoelectric focusing (IEF) also indicated that the purified CRPs differed from each other. Glycosylation was demonstrated in these purified CRPs by Digoxigenin kits, neuraminidase treatment and binding with lectins. The presence of N-linked sugar moiety was confirmed by N-glycosidase F digestion. The presence of sialic acid, glucose, galactose and mannose has been demonstrated by gas liquid chromatography, mass spectroscopic and fluorimetric analysis. Matrix-assisted laser-desorption ionization analysis of the tryptic digests of three CRPs showed systematic absence of two peptide fragments, one at the N-terminus and the other near the C-terminus. Model-building suggested that the loss of these fragments exposed two potential glycosylation sites on a cleft floor keeping the protein–protein interactions in pentraxins and calcium-dependent phosphorylcholine-binding qualitatively unaffected. Thus we have convincingly demonstrated that human CRP is glycosylated in some pathological conditions.
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July 2003
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Research Article|
July 15 2003
Induction of glycosylation in human C-reactive protein under different pathological conditions
Tanusree DAS;
Tanusree DAS
∗Immunobiology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India
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Asish SEN;
Asish SEN
∗Immunobiology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India
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Tore KEMPF;
Tore KEMPF
†Protein Analysis Facility, German Cancer Research Center, INF280, 69120 Heidelberg, Germany
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Sumit R. PRAMANIK;
Sumit R. PRAMANIK
∗Immunobiology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India
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Chhabinath MANDAL;
Chhabinath MANDAL
‡Drug Design, Development and Molecular Modelling Divisions, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India
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Chitra MANDAL
Chitra MANDAL
1
∗Immunobiology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India
1To whom correspondence should be addressed (e-mail cmandal@iicb.res.in or Chitra_mandal@yahoo.com).
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Biochem J (2003) 373 (2): 345–355.
Article history
Received:
October 31 2002
Revision Received:
February 26 2003
Accepted:
April 14 2003
Accepted Manuscript online:
April 14 2003
Connected Content
A correction has been published:
Induction of glycosylation in human C-reactive protein under different pathological conditions
Citation
Tanusree DAS, Asish SEN, Tore KEMPF, Sumit R. PRAMANIK, Chhabinath MANDAL, Chitra MANDAL; Induction of glycosylation in human C-reactive protein under different pathological conditions. Biochem J 15 July 2003; 373 (2): 345–355. doi: https://doi.org/10.1042/bj20021701
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