Synthesis and characterization of photoactivatable peptide agonists of the human thrombin receptor

FEBS Letters

Bischoff, Rainer¹  Jaquinod, Michel³  Pavirani, Andrea¹  Chong, Pele C.S.⁴  Gachet, Christian²  Rasmussen, Ulla B.¹  Tripet, Brian⁴  Cordier, Yves¹  Schlesinger, Yasmin¹ 

[1] Transgene S.A., 11 rue de Molsheim, 67082 Strasbourg Cedex, France;Centre Régional de Transfusion Sanguine, Institut National de la Santé et de la Recherche Médicale U-311, 67085 Strasbourg Cedex, France;Université Louis Pasteur, Laboratoire de Spectrometrie de Mass Bioorganique, URA 31, CNRS, 5 rue Blaise Pascal, 67008 Strasbourg Cedex, France;Connaught Laboratories Ltd., 1755 Steeles Avenue West, Willowdale, Ont., Canada

关键词: Thrombin receptor;    Photoaffinity labelling;    p-Azidophenylalanine;    Biotinylation;    Ca2+ mobilization;    HEL;    human erythroleukemia;    Fmoc;    N α-9-fluorenylmethyloxycarbonyl;    Boc;    N α-tert-butyloxycarbonyl;    RP-HPLC;    reverse-phase high-performance liquid chromatography;    ESMS;    electrospray mass spectrometry;    BSA;    bovine serum albumin;    PBS;    phosphate- buffered saline;    ELISA;    enzyme-linked immunosorbent assay;    HEPES;    (N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulf onic acid]);   

DOI  :  10.1016/0014-5793(93)00692-B

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Chemical synthesis and biochemical analysis of modified agonist peptides of the human thrombin receptor derived from the sequence SFLLRNP containing photoactivatable azido groups and biotin for sensitive detection is described. Substitution of leucine in position three with p-azidophenylalanine and extension of the C-terminus with a KGGK spacer containing biotin covalently linked to the side chain of the C-terminal lysine residue resulted in an active receptor agonist as determined by intracellular Ca²⁺ mobilization in human erythroleukemia (HEL) cells. In contrast, substitution of phenylalanine in position two with p-azidophenylalanine reduced agonist activity significantly.

【 授权许可】

Unknown