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Synthetic conformational antigen which simulates the extracellular part of the M2-muscarinic receptor: interaction with blood sera of patients suffering from idiopathic arrhythmias

Abstract

Fragments corresponding to the 83–98 sequence of the first extracellular loop and to the 168–192 and 171–182 sequences of the second extracellular loop of the M2-muscarinic receptor (antibodies to this receptor could be markers of early symptoms of heart disorders) were synthesized by solid phase method using the Fmoc-SPPS strategy. A new conformational antigen with the natural location of the disulfide bridge was prepared by selective formation of disulfide bond between the corresponding cysteine residues in the synthe-sized peptides and characterized. The comparative analysis of reactivity of the synthesized peptides towards sera from patients which had no organic heart disease was performed. A new conformational antigen was effectively bound to the sera from patients with idiopathic arrhythmias, but without symptoms of organic heart disease.

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Abbreviations

Boc:

tert-butyloxycarbonyl

BSA:

bovine serum albumin

But :

tert-butyl

DCM:

dichloromethane

DIPEA:

N,N-diisopropylethylamine

Fmoc:

9-fluorenylmethyloxycar-bonyl

HOBT:

1-hydroxybenzotriazole

MALDI:

matrix-assisted laser desorption/ionization

NMP:

N-methylpyrroli-done

MePip:

4-methylpiperidine

PyS:

2-pyridinesulfenyl

ODS:

octadecylsilyl

Pmc:

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

TBTU:

N,N,N,N′-tetramethyl-O-(benzotriazole-1-yl)uronium tetrafluoroborate

TIBS:

triisobutyl silane

TFA:

tri-fluoroacetic acid

HPLC:

high performance liquid chromatog-raphy

EIA:

enzyme immunoassay

CRD:

cardiac rhythm disturbance

SPPS:

solid phase peptide synthesis. Other abbreviations used were recommended by IUPAC-IUB

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Correspondence to M. E. Palkeeva.

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Original Russian Text © M.E. Palkeeva, M.V. Sidorova, A.S. Molokoedov, A.A. Az’muko, Zh.D. Bespalova, T.V. Sharf, E.N. Mamochkina, E.E. Efremov, M.M. Rogova, N.A. Mironova, K.A. Zykov, S.P. Golitsyn, 2013, published in Bioorganicheskaya Khimiya, 2013, Vol. 39, No. 3, pp. 285–292.

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Palkeeva, M.E., Sidorova, M.V., Molokoedov, A.S. et al. Synthetic conformational antigen which simulates the extracellular part of the M2-muscarinic receptor: interaction with blood sera of patients suffering from idiopathic arrhythmias. Russ J Bioorg Chem 39, 252–258 (2013). https://doi.org/10.1134/S1068162013030102

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  • DOI: https://doi.org/10.1134/S1068162013030102

Keywords

  • idiopathic arrhythmia
  • antibodies to M2-muscarinic receptor
  • conformational peptide antigens
  • synthetic peptides
  • selective disulfide bond formation
  • solid phase synthesis