Molecular forms of the putative enkephalin precursor BAM-12P in bovine adrenal, pituitary, and hypothalamus.
Molecular forms of the putative enkephalin precursor BAM-12P in bovine adrenal, pituitary, and hypothalamus.
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牛肾上腺、垂体和下丘脑中假定的脑啡肽前体 BAM-12P 的分子形式。
DOI：
10.1073/pnas.79.6.2023
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发表时间：
1982
影响因子：
11.1
通讯作者：
Guillemin,R
中科院分区：
文献类型：
--
作者：
Baird,A;Ling,N;Böhlen,P;Benoit,R;Klepper,R;Guillemin,R
一种高度特异性的放射免疫测定方法已经被开发出来，用于检测一种假定的肾上腺髓质[Met]脑啡肽前体BAM-12P （tyr - gly - gly - ph -Met- arg - arg - val - gly - arg - pro - glu- - -）。bam12p抗体识别从Arg7到Glu12的肽的cooh末端片段，不会与[Met5]-或[Leu5]脑啡肽或它们的任何cooh末端赖氨酸或精氨酸延伸类似物发生交叉反应。与BAM-12P5-12的交叉反应性为100%，与BAM-12P5-12酰胺的交叉反应性为0.3%，表明cooh末端Glu-OH的特异性。利用这些抗体，我们在牛肾上腺髓质提取物中测量了三种形式的bam12p样免疫反应性，其中主要形式（大于90%）的分子量与bam12p相对应。牛肾上腺皮质提取物含有1%的bam12p样物质（Mr约为1400，每个腺体20 ng），可能是由于与肾上腺髓组织交叉污染。牛脑垂体神经中间体提取物中bam12p样物质的主要形式比真实的bam12p分子量更高（Mr约为4000）；剩余材料（10%）按分子量对应于BAM-12P。牛垂体前叶的粗提取物或纯化提取物中没有检测到bam12p样免疫反应性。牛下丘脑组织的提取物含有少量的BAM-12P免疫反应性（每个片段约2 ng），可以作为一种分子形式检测到对应于1400道尔顿分子。结果表明，在肾上腺髓质中发现的脑啡肽前体也可能存在于垂体和下丘脑中。此外，这种分子的加工似乎是组织特异性的。
A highly specific radioimmunoassay for one of the putative adrenomedullary [Met]enkephalin precursors, BAM-12P (Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg-Pro-Glu-OH), has been developed. The BAM-12P antibodies recognize the COOH-terminal fragment of the peptide from Arg7 to Glu12 and do not crossreact with [Met5]- or [Leu5]enkephalin or any of their COOH-terminal lysine or arginine extended analogs. Specificity for the COOH-terminal Glu-OH is suggested by the 100% crossreactivity with BAM-12P5-12 and 0.3% crossreactivity with BAM-12P5-12 amide. Using these antibodies, we have measured three forms of BAM-12P-like immunoreactivity in extracts of bovine adrenal medulla, of which the major form (greater than 90%) corresponds to BAM-12P by molecular weight. Extracts of bovine adrenal cortex contain 1% the amount of a BAM-12P-like material (Mr approximately 1400; 20 ng per gland), possibly due to crosscontamination with adrenomedullary tissue. The major form of BAM-12P-like material in extracts of bovine neurointermediate pituitaries is of higher molecular weight than authentic BAM-12P (Mr approximately 4000); the remaining material (10%) corresponds to BAM-12P by molecular weight. There is no detectable BAM-12P-like immunoreactivity in crude or purified extracts of bovine anterior pituitaries. Extracts of bovine hypothalamic tissues contain small amounts of BAM-12P immunoreactivity (approximately 2 ng per fragment) which can be detected as one molecular form corresponding to a 1400-dalton molecule. The results indicate that the enkephalin precursor found in the adrenal medulla also may be present in the pituitary and hypothalamus. Furthermore, the processing of this molecule appears to be tissue-specific.