Preparation of a photoaffinity probe for the Man6P/IGF-II receptor.
A photoaffinity probe, 2-(4-phosphopentamanniminophenyl)ethyl-(4-azido) salicylamide, was prepared for photolabelling of the mannose 6-phosphate/insulin-like growth factor II receptor. The probe was synthesized from phosphopentamannan and 2-(4-aminophenyl)ethylamine followed by coupling with photosensitive 4-azidosalicylic acid. The 125I-labelled probe was reacted with receptor purified from rat liver under irradiation with ultraviolet light. Fluorography of the reaction product on a polyacrylamide gel effectively detected the receptor coupled to the photoprobe as a molecular mass of 250 kDa. Specific binding of the probe or of radiolabelled insulin-like growth factor II was not inhibited by the growth factor or by mannose 6-phosphate, confirming different binding sites to the receptor between the ligands.
Miyagishima
T
T
Biochemistry Laboratory, Hokkaido University School of Medicine, Sapporo, Japan.
Gasa
S
S
Honke
K
K
Miyazaki
T
T
Makita
A
A
D000345:Affinity Labels / Q000138:chemical synthesis* / Q000737:chemistry; D000818:Animals; D001386:Azides / Q000138:chemical synthesis* / Q000737:chemistry; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D007335:Insulin-Like Growth Factor II / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008351:Mannans / Q000138:chemical synthesis* / Q000737:chemistry; D008360:Mannosephosphates / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D017451:Receptors, Somatomedin / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D003432:Cross-Linking Reagents; D008351:Mannans; D008360:Mannosephosphates; D017451:Receptors, Somatomedin; C082173:2-((4-phosphopentamanniminophenyl)ethyl)-4-azidosalicylamide; C027693:mannose-6-phosphate; D007335:Insulin-Like Growth Factor II