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成年及发育中的非洲爪蟾中的烯醇化酶同工酶以及一个克隆的烯醇化酶序列的特征分析

Enolase isoenzymes in adult and developing Xenopus laevis and characterization of a cloned enolase sequence.

作者信息

Segil N, Shrutkowski A, Dworkin M B, Dworkin-Rastl E

机构信息

Department of Biological Sciences, Sherman Fairchild Center, Columbia University, New York, NY 10027.

出版信息

Biochem J. 1988 Apr 1;251(1):31-9. doi: 10.1042/bj2510031.

DOI:10.1042/bj2510031
PMID:3390159
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1148960/
Abstract

As part of a study of glycolysis during early development we have examined the pattern of expression of enolase isoenzymes in Xenopus laevis. In addition, the nucleotide sequence of a cDNA clone coding for the complete amino acid sequence of one enolase gene (ENO1) in X. laevis was determined. X. laevis ENO1 shows highest homology to mammalian non-neuronal enolase. Analysis of enolase isoenzymes in X. laevis by non-denaturing electrophoresis on cellulose acetate strips revealed five isoenzymes. One form was present in all tissues tested, two additional forms were expressed in oocytes, embryos, adult liver and adult brain, and two further forms were restricted to larval and adult muscle. Since enolase is a dimer, three different monomers (gene products) could account for the observed number of isoenzymes. This pattern of enolase isoenzyme expression in X. laevis differs from that of birds and mammals. In birds and mammals the most acidic form is neuron-specific and there is only one major isoenzyme expressed in the liver. RNAase protection experiments showed the presence of ENO1 mRNA in oocytes, liver and muscle, suggesting that it codes for a non-tissue-restricted isoenzyme. ENO1 mRNA concentrations are high in early oocytes, decrease during oogenesis and decrease further after fertilization. Enolase protein, however, is maintained at high concentrations throughout this period.

摘要

作为早期发育过程中糖酵解研究的一部分，我们检测了非洲爪蟾烯醇化酶同工酶的表达模式。此外，还测定了编码非洲爪蟾一个烯醇化酶基因（ENO1）完整氨基酸序列的cDNA克隆的核苷酸序列。非洲爪蟾ENO1与哺乳动物非神经元烯醇化酶的同源性最高。通过在醋酸纤维素条上进行非变性电泳分析非洲爪蟾的烯醇化酶同工酶，发现了五种同工酶。有一种形式存在于所有测试组织中，另外两种形式在卵母细胞、胚胎、成年肝脏和成年大脑中表达，还有两种形式仅存在于幼虫和成年肌肉中。由于烯醇化酶是一种二聚体，三种不同的单体（基因产物）可以解释观察到的同工酶数量。非洲爪蟾烯醇化酶同工酶的这种表达模式与鸟类和哺乳动物不同。在鸟类和哺乳动物中，最酸性的形式是神经元特异性的，肝脏中只表达一种主要的同工酶。核糖核酸酶保护实验表明，卵母细胞、肝脏和肌肉中存在ENO1 mRNA，这表明它编码一种非组织限制性同工酶。ENO1 mRNA浓度在早期卵母细胞中较高，在卵子发生过程中降低，受精后进一步降低。然而，在整个这一时期，烯醇化酶蛋白的浓度保持在较高水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/f379821fac70/biochemj00234-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/79fe7c648d1b/biochemj00234-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/6d418c0eee15/biochemj00234-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/f379821fac70/biochemj00234-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/79fe7c648d1b/biochemj00234-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/6d418c0eee15/biochemj00234-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ceb5/1148960/f379821fac70/biochemj00234-0046-a.jpg

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