Synthèse d'une sonde biotinylée à bras clivable allongé pour l'isolement des récepteurs de l'angiotensine II

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Abstract
Synthesis of a Biotinylated Probe with an Extended Cleavable Arm for Angiotensin II Receptors PurificationWe have synthesized a new biotinylated probe for angiotensin II receptors studies: biotinyl‐NH(CH2)2SS(CH2)2CO‐Gly‐ϵ Ahx‐[Ala1, Phe(4N3)8]angiotensin II (5). This molecule can be photoactivated through an arylazido group. 1H‐NMR studies suggest that it adopts an extended conformation which should allow a simultaneous recognition of both streptavidin and hormone receptor. It has a good affinity for receptors (Kd = 1 nM) and hence is a promising tool in their detection (autoradiography, gold‐, ferritin‐, enzyme‐, or fluorescent streptavidin derivatives) and separation (cell sorting, affinity chromatography). It can be monoiodinated (°6) at its tyrosine residue without a significant loss of affinity. Its extended cleavable arm allows an easy recovery of the ‘probe‐receptor’ complex from streptavidin. An HPLC monitoring of the synthesis is described, particularly of the segment coupling 1 + 2 in presence of (benzotriazol‐1‐yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP). This method can be used as well for synthesis of the D‐Phe8 derivative that has antagonist properties.