Approaches to radioiodination of insect neuropeptides
Approaches to radioiodination of insect neuropeptides
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DOI：
10.1016/s0196-9781(02)00192-4
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发表时间：
2002-11-01
期刊：
影响因子：
3
通讯作者：
Brown, MR
中科院分区：
文献类型：
--
作者：
Crim, JW;Garczynski, SF;Brown, MR
高质量的放射性碘标记神经肽是放射免疫分析和受体结合分析的基础。比较了用~（125）I（I-125）直接和间接标记神经肽的方法。使用配备有在线伽马检测器和UV吸光度检测器的HPLC来评价所选择的标记方法和产品。用氯胺-T处理[Y-1]-adipokineticase-I（[Y-1]-AKH-I）引起氧化损伤，而在H2 O2存在下用乳过氧化物酶进行酶标记产生了良好的完整产率，显然是单碘化肽。用氯胺-T标记FMRFamide相关肽（YGGFMRFa）显然形成甲硫氨酸亚砜，其随后可以用二硫苏糖醇还原。通常可获得高比活性的产物。(C)2002年爱思唯尔科技有限公司All rights reserved.
High quality radioiodinated neuropeptides are essential to radioimmunoassays (RIA) and receptor binding assays. Approaches of direct and indirect labeling of neuropeptides with (125)Iodine (I-125) are compared. An HPLC equipped with an in-line gamma detector and UV absorbance detector was used to evaluate selected labeling methods and products. Treatment of [Y-1]-adipokinetic hormone-I ([Y-1]-AKH-I) with chloramine-T caused oxidative damage, whereas enzymatic labeling with lactoperoxidase in the presence of H2O2 produced a good yield of intact, apparently monoiodinated peptide. Labeling of the FMRFamide-related peptide (YGGFMRFa), with chloramine-T apparently formed the methionine sulfoxide, which subsequently could be reduced with dithiothreitol. Products of high specific activity typically are achievable. (C) 2002 Elsevier Science Inc. All rights reserved.