Purification and sequence of rat oxyntomodulin.
Purification and sequence of rat oxyntomodulin.
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大鼠泌酸调节素的纯化和序列。
DOI：
10.1073/pnas.91.20.9362
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发表时间：
1994
影响因子：
11.1
通讯作者：
ReeveJr,JR
中科院分区：
文献类型：
--
作者：
Collie,NL;Walsh,JH;Wong,HC;Shively,JE;Davis,MT;Lee,TD;ReeveJr,JR
关于大鼠肠胰高血糖素（含有胰高血糖素序列的肠肽）的结构信息先前已基于cDNA、免疫学和色谱数据。我们的兴趣是测试合成的肠胰高血糖素肽在大鼠中的生理作用，这要求我们精确地识别体内存在的形式。从胰高血糖素原基因序列的知识，我们合成了一个肠胰高血糖素C-末端八肽共同提出的肠胰高血糖素形式，胰高血糖素和胃泌酸调节素，但没有共享与胰高血糖素重叠的序列。然后，我们开发了一种放射免疫测定法，使用针对肠胰高血糖素肽特异性的八肽抗体，而不与胰高血糖素发生交叉反应。提取大鼠肠，并通过以下肠胰高血糖素C-末端八肽样免疫反应性通过几个HPLC纯化步骤纯化一种假定的肠胰高血糖素形式。通过氨基酸组成、微序列和质谱分析对材料进行结构表征，将肽鉴定为大鼠胃泌酸调节素。该37个残基的肽由胰高血糖素加上C-末端延伸Lys-Arg-Asn-Arg-Asn-Asn-Ile-Ala组成。这现在允许合成用于生理学研究的内源性大鼠胃泌酸调节素的明确重复。
Structural information about rat enteroglucagon, intestinal peptides containing the pancreatic glucagon sequence, has been based previously on cDNA, immunologic, and chromatographic data. Our interests in testing the physiological actions of synthetic enteroglucagon peptides in rats required that we identify precisely the forms present in vivo. From knowledge of the proglucagon gene sequence, we synthesized an enteroglucagon C-terminal octapeptide common to both proposed enteroglucagon forms, glicentin and oxyntomodulin, but sharing no sequence overlap with glucagon. We then developed a radioimmunoassay using antibodies raised against the octapeptide that was specific for enteroglucagon peptides without cross-reacting with glucagon. Rat intestine was extracted, and one presumptive enteroglucagon form was purified by following the enteroglucagon C-terminal octapeptide-like immunoreactivity through several HPLC purification steps. Structural characterization of the material by amino acid composition, microsequence, and mass spectral analyses identified the peptide as rat oxyntomodulin. The 37-residue peptide consists of pancreatic glucagon plus the C-terminal extension, Lys-Arg-Asn-Arg-Asn-Asn-Ile-Ala. This now permits synthesis of an unambiguous duplicate of endogenous rat oxyntomodulin for physiological studies.