从固氮菌2-oxoglutarate脱氢酶复杂vinelandii。2。基因的分子克隆和序列分析编码succinyltransferase组件。
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Westphal啊,德角
从固氮菌2-oxoglutarate脱氢酶复杂vinelandii。2。基因的分子克隆和序列分析编码succinyltransferase组件。
欧元。1990 1月12日,187 (1):235 - 9。
- PubMed ID
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2404760 (在PubMed]
- 文摘
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核苷酸序列编码succinyltransferase组件(E2o) 2-oxoglutarate脱氢酶复杂的固氮菌vinelandii已经确定。以前在大肠杆菌克隆的基因编码lipoamide脱氢酶从a vinelandii报道[Westphal A.H. &德角,a(1988)欧元。j。172, 299 - 305]。3.2 kb片段用于序列测定的主要部分包含基因编码succinyltransferase。完整的E2o基因以及2-oxoglutarate脱氢酶的基因编码组件,居住在一个14.7 kb片段的3.2 kb subcloned片段。蛋白质编码序列的基因由1200个基点(400密码子,包括8月起始密码子和佐治亚大学终止密码子)。它是分开2-oxoglutarate脱氢酶的基因编码组件由42个基点。没有发现大肠coli-like启动子序列。一个假定的ribosome-binding站点位于15英国石油公司上游从一开始的密码子。没有发现终结者序列下游的终止密码子。 This makes it likely that the three genes of the oxoglutarate dehydrogenase complex are transcribed as a single mRNA transcript analogous to the pyruvate dehydrogenase complex in E. coli. The intact gene was subcloned from the 14.7-kb fragment and brought to high expression under the influence of the vector-encoded lacZ promoter. The similarity with the E. coli enzyme is high with 63% identity. Like the enzyme from E. coli, it consists of a single lipoyl-binding domain, a putative E1- and E3-binding domain and a catalytic domain. The main difference is found in a 31-residue sequence rich in alanine and proline located between the lipoyl domain and the putative E1- and E3-binding domain. This sequence, usually found in acetyltransferases and there identified as a highly mobile region by 1H-NMR, is replaced by a more polar, charged region in the E. coli enzyme.