Preparation of [125I-Tyr27,Leu5]beta h-endorphin and its use for crosslinking of opioid binding sites in human striatum and NG108-15 neuroblastoma-glioma cells.

Abstract

A radioligand suitable for crosslinking studies to opioid receptors has been obtained by radioiodination and purification of the monoiodotyrosine-27 derivative of the synthetic human beta-endorphin (beta h-endorphin) analogue [5-leucine]beta h-endorphin. The derivative, [27-[125I]monoiodotyrosine,5-leucine]beta h-endorphin, was crosslinked to human striatal (caudate and putamen) and NG108-15 neuroblastoma-glioma cell membranes by using disuccinimidyl suberate. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis under reducing conditions revealed four specifically labeled bands at 68, 40, 30, and 25 kDa for both human caudate and putamen, whereas NG108-15 cell membranes gave specifically labeled bands at 92, 56, 38, and 23 kDa.

• Authors: D M Helmeste, R G Hammonds Jr, C H Li
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 83 Issue 13 Pg. 4622-5 (Jul 1986) ISSN: 0027-8424 [Print] United States
• PMID: 3014499 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Cross-Linking Reagents
• Endorphins
• Receptors, Opioid

Topics

• Adult
• Animals
• Caudate Nucleus (metabolism)
• Cell Line
• Cell Membrane (metabolism)
• Cross-Linking Reagents
• Endorphins (chemical synthesis, metabolism)
• Glioma (metabolism)
• Humans
• Male
• Mice
• Molecular Weight
• Neuroblastoma (metabolism)
• Putamen (metabolism)
• Rats
• Receptors, Opioid (metabolism)