Preparation of tritium labeled beta-endorphin suitable for studying brain receptor interactions |
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Authors: | R G Hammonds N Ling D Puett |
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Institution: | 1. Department of Biochemistry, Vanderbilt University Nashville, TN 37232 USA;2. Neuroendocrinology Laboratory, The Salk Institute La Jolla, CA 92037 USA |
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Abstract: | The opioid peptide (porcine) β-endorphin has been tritiated using reductive methylation to prepare a derivative containing mainly 3H]dimethyllysine. The tritiated β-endorphin has a specific activity of 9.8 Ci/mmol and is stable for an extended period of time. The labeled peptide binds reversibly to rat brain membrane preparations with a dissociation constant of 0.4 ± 0.1 nM and a receptor content of 23 ± 2 pmol/g brain. Under the conditions used, there is evidence for only one class of receptors. The technique employed for tritium labeling of β-endorphin should also be applicable to various other peptides including α-endorphin, γ-endorphin, and C′-fragment that have been found in brain and pituitary. |
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Keywords: | Me methylated lysine SDS sodium dodecyl sulfate |
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