Binding affinities to rat brain synaptosomes – synthesis of biotinylated analogues of Substance P

Photoactivatable substance P (SP) derivatives containing the p‐benzoylbenzoic moiety at the N‐terminal α‐amino group of Arg 1 or at the ε‐amino group of Lys 3 were prepared. Both derivatives also had a p‐hydroxyphenylpropionyl group for radioiodination. To obtain the analogue with the photolabel at Arg 1, SP was first reacted with N‐hydroxysuccinimide p‐hydroxyphenylpropionate, the Lys 3‐modified derivative was isolated by reversed‐phase high‐performance liquid chromatography (HPLC), reacted with N‐hydroxysuccinimide p‐benzoylbenzoate and purified by HPLC. To place the photolabel at Lys 3, the order of the reactions was reversed. The structure of the derivatives obtained was confirmed by mass spectrometry. The interaction of the derivatives obtained and of their 125I‐labeled forms with the NK‐1 neurokinin receptor from the rat brain, as well as with the nicotinic acetylcholine receptor from Torpedo electrocytes was analyzed. The results obtained supported by the data from the literature indicate that benzoylbenzoic acid derivatives should not be considered as universal photolabels, which ensure in all cases a high level of photo‐cross‐linking.

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“…Previous structure-activity studies (24,25) on SP have shown that modification of the Lys 3 side-chain has no…”

Section: Resultsmentioning

confidence: 94%

Substance P derivatives with photoactivatable labels in the N‐terminal part of the molecule

Kasheverov

Utkin

Franke

et al. 1997

The Journal of Peptide Research

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

Seyer

Aumelas

Marie

et al. 1989

Helvetica Chimica Acta

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We have synthesized a new biotinylated probe for angiotensin I1 receptors studies: biotinyl-NH(CH,),-SS-(CH2)2-CO-Gly-cAhx-[Ala', Phe(4N3)']angiotensin I1 (5). This molecule can be photoactivated through an arylazido group. 'H-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 ( 4 ) 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-l-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP). This method can be used as well for synthesis of the D-Phe' derivative that has antagonist properties.

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Solid-phase synthesis and applications of N-(S-acetylmercaptoacetyl) peptides

Drijfhout

Bloemhoff

Poolman

et al. 1990

Analytical Biochemistry

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Binding affinities to rat brain synaptosomes – synthesis of biotinylated analogues of Substance P

Cited by 11 publications

References 21 publications

Substance P derivatives with photoactivatable labels in the N‐terminal part of the molecule

Substance P derivatives with photoactivatable labels in the N‐terminal part of the molecule

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

Solid-phase synthesis and applications of N-(S-acetylmercaptoacetyl) peptides

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