Synthesis of a biotinylated, iodinatable, and photoactivatable probe for angiotensin receptors.
We propose here a biotinyl-aminohexanoyl-[Ala1, Phe(4N3)8]angiotensin II analog as a radioiodinatable and photoactivatable probe for covalent labeling, detection and isolation of angiotensin receptors. A combination of solid phase and minimum-protection segment-coupling strategy using hexafluorophosphate of (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium (BOP) as a coupling reagent is proposed for the synthesis of this probe. Optimized yields were obtained by HPLC monitoring of all reactions. A complete n.m.r. study suggests an extended conformation of this molecule, allowing a simultaneous recognition of receptor and avidin. The probe binds with high affinity (Kd = 2 nM) to angiotensin II receptors from rat liver membranes.
Seyer
R
R
CNRS-INSERM Center for Pharmacology-Endocrinology, Montpellier, France.
Aumelas
A
A
Tence
M
M
Marie
J
J
Bonnafous
J C
JC
Jard
S
S
Castro
B
B
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors; D057911:Angiotensin Receptor Antagonists; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008027:Light; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015335:Molecular Probes; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011945:Receptors, Angiotensin / Q000378:metabolism / Q000502:physiology*; D013056:Spectrophotometry, Ultraviolet
D057911:Angiotensin Receptor Antagonists; D007457:Iodine Radioisotopes; D015335:Molecular Probes; D011945:Receptors, Angiotensin; D000804:Angiotensin II; C061954:angiotensin II, Sar(1)-Phe(8)-; C061953:biotinyl-epsilon-aminohexanoyl-alanyl-arginyl-valyl-tyrosyl-isoleucyl-histidyl-prolyl-phenylalanyl(4N3)-hydroxy; D001710:Biotin