Cloning and functional analysis of C. elegans 7B2.
Cloning and functional analysis of C. elegans 7B2.
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线虫 7B2 的克隆和功能分析。
DOI：
10.1089/dna.1998.17.727
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发表时间：
1998
影响因子：
3.1
通讯作者：
Dickerson,IM
中科院分区：
文献类型：
--
作者：
Lindberg,I;Tu,B;Muller,L;Dickerson,IM
神经内分泌蛋白7 B2是激素原转化酶2（PC 2）的结合蛋白，是proPC 2在细胞内转化为活性PC 2所必需的。全长7 B2及其羧基末端31个残基肽（CT肽）都能够有效抑制PC 2;因此，7 B2蛋白调节PC 2的生物合成和活性。脊椎动物7 B2是高度保守的（92%-97%同源性），因此，物种比较在评估负责生物活性的关键蛋白质结构域方面没有提供信息。本文报道了秀丽隐杆线虫7 B2蛋白的克隆。虽然与脊椎动物序列弱保守（与小鼠7 B2的相似性为23%），但秀丽隐杆线虫7 B2含有特征性PPNPCP基序以及CT肽内的高度保守七肽。体外测定，C. elegans 7 B2对重组脊椎动物PC 2具有显著的抑制活性（IC 50130 nM），在两个功能测试中，C. elegans 7 B2促进proPC 2的激活。我们的结论是，尽管氨基酸保守性总体较低，但7 B2内的两个功能结构域在C. elegans和脊椎动物蛋白质。
The neuroendocrine protein 7B2 is a binding protein for the prohormone convertase 2 (PC2) and is required for the intracellular conversion of proPC2 to active PC2. Both full-length 7B2 and its carboxy-terminal 31-residue peptide (CT peptide) are capable of potent inhibition of PC2; the 7B2 protein thus regulates both the biosynthesis and the activity of PC2. Vertebrate 7B2s are highly conserved (92%-97% homology), and thus, species comparison has not been informative in assessing the crucial protein domains responsible for bioactivity. We here report the cloning of theCaenorhabditis elegans7B2 protein. Although weakly conserved with the vertebrate sequences (23% similarity with mouse 7B2), C.elegans 7B2 contains the signature PPNPCP motif as well as a highly conserved heptapeptide within the CT peptide. Inin vitroassays,C. elegans7B2 possessed significant inhibitory activity against recombinant vertebrate PC2 (IC50130 nM), and in two functional tests, the amino-terminal domain ofC. elegans7B2 facilitated the activation of proPC2. We conclude that despite low amino acid conservation overall, both functional domains within 7B2 have been conserved between theC. elegansand the vertebrate proteins.