Elucidation of a minimal immunoreactive site of vertebrate calmodulin.

Elucidation of a minimal immunoreactive site of vertebrate calmodulin.
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脊椎动物钙调蛋白最小免疫反应位点的阐明。

DOI:
10.1016/0003-9861(83)90481-2
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发表时间:
1983
影响因子:
3.9
通讯作者:
Erickson,BW
Erickson,BW
中科院分区:
生物学3区
文献类型:
--
作者:
VanEldik,LJ;Watterson,DM;Fok,KF;Erickson,BW

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对应于脊椎动物钙调蛋白第137-143位残基的七肽Asn Tyr Glu Phe瓦尔Gln NH 2与整个148个残基的蛋白质具有同样的免疫反应性。先前已经描述了用于产生抗脊椎动物钙调蛋白的抗血清的可重复的和快速的方法(L. J.货车Eldik和D. M. Watterson(1981)J.Biol.Chem.256,4205-4210)。大多数的抗血清引起的这种方法与脊椎动物钙调素的COOH-末端结构域的主要免疫反应区域(残基127-144)。在这份报告中,钙调素的最小部分所需的反应性与抗血清,很容易区分各种类型的钙调素的定义。这些研究表明,在放射免疫分析中,七个氨基酸残基的线性段显示类似于完整钙调素的竞争曲线的竞争曲线。该七肽是最小的钙调素片段,也是135-145区域中唯一与抗钙调素血清显示定量免疫反应性的七残基片段。这些数据表明,这七肽是钙调蛋白的主要免疫反应位点。然而,当这个免疫反应位点七肽与载体结合并注射到兔子体内时,它不会引发与天然蛋白质反应的抗血清。这些研究表明,在动物中产生的抗血清的定量免疫反应性可以在小的肽段中发现,并且对于钙调蛋白,与天然蛋白质分子反应的抗肽抗体的产生的要求并不像肽区域的表面暴露那样简单。
The heptapeptide AsnTyrGluGluPheValGlnNH2corresponding to residues 137–143 of vertebrate calmodulin is as immunoreactive as the entire 148-residue protein. A reproducible and rapid procedure for producing antisera against vertebrate calmodulin has been previously described (L. J. Van Eldik and D. M. Watterson (1981)J. Biol. Chem.256, 4205–4210). Most of the antisera elicited by this method react with a major immunoreactive region (residues 127–144) in the COOH-terminal domain of vertebrate calmodulin. In this report, the minimum segment of calmodulin required for reactivity with an antiserum that readily distinguishes various types of calmodulins is defined. These studies demonstrate that a linear segment of seven amino acid residues shows a competition curve in radioimmunoassay resembling the competition curve of intact calmodulin. This heptapeptide is the smallest calmodulin segment and the only sevenresidue segment in the 135–145 region that shows quantitative immunoreactivity with the anti-calmodulin serum. These data demonstrate that this heptapeptide is a major immunoreactive site of calmodulin. However, when this immunoreactive site heptapeptide is conjugated to a carrier and injected into rabbits, it does not elicit antisera that react with the native protein. These studies demonstrate that quantitative immunoreactivity of antisera produced in animals can be found in small peptide segments and that, for calmodulin, the requirements for production of anti-peptide antibodies that react with the native protein molecule are not as simple as surface exposure of the peptide region.
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发表时间: 1982-02
期刊: The Journal of biological chemistry
影响因子: --
作者:
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