Purification and characterization of neurotensin receptor from rat brain with special reference to comparison between newborn and adult age rats. |
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Authors: | Y Miyamoto-Lee S Shiosaka M Tohyama |
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Institution: | Department of Neuroanatomy, Osaka University Medical School, Japan. |
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Abstract: | Scatchard analysis of saturation curves was performed to compared newborn and adult rat neurotensin receptor using 3H] neurotensin as a tracer. The membrane fraction of newborn rat cerebral cortex has a single population of neurotensin receptor (Kd = 0.13 nM, Bmax = 710 fmol/mg protein), whereas adults have two distinct neurotensin binding sites (high affinity site, Kd1 = 0.13 nM; low affinity site, Kd2 = 20 nM). High affinity neurotensin receptor, solubilized with digitonin, was purified from newborn rat cortex by affinity chromatography. An overall purification of 14,000-fold was achieved. The binding of 3H] neurotensin to the purified receptor is saturable and specific, with a Kd of 0.45 nM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the presence of 2-mercaptoethanol revealed purified material of a single major band of Mr = 55,000. |
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