Characterization of the adipokinetic hormone receptor from the fat body of Manduca sexta

Regulatory Peptides Pub Date : 1995-06-27 DOI:10.1016/0167-0115(95)00046-E

Rolf Ziegler , Ronald D. Jasensky , Hiromi Morimoto

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引用次数: 57

Abstract

A tritium labeled Manduca sexta adipokinetic hormone (M-AKH) was synthesized (pE-L-T-[p³H]F-T-S-S-W-G-NH₂) (specific activity 27 Ci/mmol) which was fully active in a bioassay. It was used in a filtration based binding assay to characterize the M-AKH receptor from the fat body of M. sexta. Membrane fractions were prepared from fat body and optimal binding conditions were determined. A K_d of 7·10⁻¹⁰ M was determined and the receptor concentration estimated to be 0.5 pmol/mg membrane protein. No receptor binding was found when membranes were prepared from brain, heart or flight muscle of M. sexta or from fat body of the cockroach Blaberus discoidalis. However, specific binding was found with membrane preparations from the pterothoracic ganglion of M. sexta. The membranes from the ganglion had a much smaller number of binding sites than the fat body membranes, however, the binding was specific and observed in each experiment.

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Manduca sexta脂肪体脂肪动力激素受体的研究

合成了一种氚标记的Manduca sexta脂肪动力学激素(M-AKH) (pE-L-T-[p3H]F-T-S-S-W-G-NH2)，其比活性为27 Ci/mmol，在生物实验中具有充分活性。它被用于基于过滤的结合实验来表征m.s sexta脂肪体的M-AKH受体。以脂肪体为原料制备膜组分，确定最佳结合条件。测定了Kd为7·10−10 M，估计受体浓度为0.5 pmol/mg膜蛋白。用性田鼠的脑、心、飞肌和盘状小蠊的脂肪体制备膜，均未发现受体结合。然而，发现与翼胸神经节膜制剂特异性结合。与脂肪体膜相比，神经节膜的结合位点数量要少得多，然而，这种结合是特异性的，并且在每次实验中都能观察到。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Regulatory Peptides 医学-内分泌学与代谢

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审稿时长

2 months

期刊介绍： Regulatory Peptides provides a medium for the rapid publication of interdisciplinary studies on the physiology and pathology of peptides of the gut, endocrine and nervous systems which regulate cell or tissue function. Articles emphasizing these objectives may be based on either fundamental or clinical observations obtained through the disciplines of morphology, cytochemistry, biochemistry, physiology, pathology, pharmacology or psychology.