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Synthesis and properties of a new conformational antigen which models an extracellular region of β1-adrenoreceptor

Abstract

Two fragments corresponding to the 125–133 and 206–218 sequences of a molecule of the β1-adrenoreceptor (autoantibodies to this protein are often found in patients with dilated cardiomyopathy) were synthesized by the solid phase method with the use of Fmoc technology. Two new conformational antigens were prepared by directed (regioselective) and undirected (spontaneous) formation of intramolecular and intermolecular disulfide bridges between the corresponding cysteine residues of the synthesized peptides. One of these antigens consisted of a mixture of disulfide isomers, and another antigen was an isomer with a natural arrangement of S-S bridges. Immunosorbents were obtained by immobilization of the synthesizes antigens on the bromocyanogen-activated sepharose and applied to the removal of autoantibodies in a β1-adrenoreceptor from the blood plasma of patients. We demonstrated that the sorbents on the basis of the conformational antigens were more effective in comparison with those containing linear peptide precursors.

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Abbreviations

Acm:

acetamidomethyl

DCMP:

dilated cardiomyopathy

DIC:

N,N′-diisopropylcarbodiimide

Fmoc:

9-fluorenylmethyloxycarbonyl

HOBt:

1-hydroxybenzotrazole

NMP:

N-methylpyrrolidone

TIBS:

triisobutylsilane

Pmc:

2,2,5,7,8-pentamethylchromane-6-sulfonyl

SPyr:

2-pyridinesulfenyl

SPPS:

solid phase peptide synthesis.

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Correspondence to M. V. Sidorova.

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Original Russian Text © M.V. Sidorova, M.E. Pal’keeva, A.S. Molokoedov, A.A. Az’muko, A.V. Sekridova, M.V. Ovchinnikov, P.A. Levashov, O.I. Afanasieva, Yu.V. Berestetskaya, M.I. Afanasieva, O.A. Razova, Zh.D. Bespalova, S.N. Pokrovskii, 2009, published in Bioorganicheskaya Khimiya, 2009, Vol. 35, No. 3, pp. 311–322.

Other abbreviations correspond to IUPAC-IUB: Eur. J. Biochem, 1994, vol. 183, pp. 9–37.

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Sidorova, M.V., Pal’keeva, M.E., Molokoedov, A.S. et al. Synthesis and properties of a new conformational antigen which models an extracellular region of β1-adrenoreceptor. Russ J Bioorg Chem 35, 285–295 (2009). https://doi.org/10.1134/S1068162009030029

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  • DOI: https://doi.org/10.1134/S1068162009030029

Key words

  • peptide antigens
  • spontaneous and regioselective formation of disulfide bonds
  • β1-adrenoreceptor
  • dilated cardiomyopathy