Analog of the fragment 197-221 of beta-1 adrenoreceptor
Analog of the fragment 197-221 of 1-adrenoreceptor
Beta-1 adrenoreceptor, Beta-1 adrenoceptor
Analog of the fragment 197-221 of 1- adrenoreceptor
The peptide was prepared by automatic solid phase synthesis (SPPS) using the Fmoc strategy starting from 0.25 mmol of commercial N Fmoc-Lys(Boc)-polymer (Bachem, Switzerland). A copolymer of styrene with 1% divinylbenzene with a hydroxymethylphenyloxymethyl anchoring group (the Wang-polymer) was used as a carrier. The side functions of the amino acids were blocked by the following protective groups: But for hydroxyl functions of Ser and Tyr; But for carboxyl functions of Asp and Glu; Boc for -amino functions of Lys; Trt for carboxylamide group of Asn, sulfhydryl functions of residues Cys198 and Cys220; Acm for sulfhydryl functions of residues Cys209 and Cys215, and Pbf for guanidine function of Arg. The amino acid chain was elongated stepwise starting from C-terminus using carbodiimide in the presence of 1-hydroxybenzotriasole. When the SPPS was finished, the peptide was cleaved from carrier with the simultaneous removal of the acid-labile protective groups by treatment with trifluoracetic acid with the addition of water, triisopropylsilane and dithiothreitole. The synthesized [Cys(Acm)2]peptide was purified by HPLC to a purity 85-90%. The intramolecular S-S bridge between Cys198 and Cys220 was with hydrogen peroxide at pH 8. The target monocyclic intermediate of peptide was purified by HPLC to a purity 90-95%. The product was characterized by MALDI-TOF mass spectrometry. The second disulfide bond between Cys209 and Cys215 was formed by the treatment of monocyclic intermediate [Cys(Acm)2]peptide with iodine (for simultaneous cleavage of Acm group and formation S-S bridge) in methanol. The bicyclic target product was purified by HPLC to a purity 98% and characterized by MALDI-TOF mass spectrometry.
AUTHORS STATE THAT THIS IS AN ANALOG OF THE FRAGMENT 197-221 OF BETA-1 ADRENORECEPTOR.
2 mM protein, 1 mM sodium azide, 20 mM [U-100% 2H] sodium acetate, 90% H2O/10% D2O
2 mM protein, 1 mM sodium azide, 20 mM [U-100% 2H] sodium acetate, 100% D2O
beta1-adrenoreceptor, modified extracellular loop 197-221, autoantibodies, cardiomyopathy, arrhythmia, IMMUNE SYSTEM