MalFGK complex assembly and transport and regulatory characteristics of MalK insertion mutants

MalFGK complex assembly and transport and regulatory characteristics of MalK insertion mutants
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MalFGK复合物组装和运输以及MalK插入突变体的调控特征

DOI:
10.1128/jb.179.4.1337-1343.1997
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发表时间:
1997
影响因子:
3.2
通讯作者:
B. Traxler
B. Traxler
中科院分区:
生物学3区
文献类型:
--
作者:
John Lippincott;B. Traxler

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MalK是一种外周细胞质膜蛋白,在大肠杆菌中具有多种活性。它与完整的细胞质膜蛋白MalF和MalG结合形成麦芽糖转运复合物(MalFGK),这是atp结合盒(ABC)超家族蛋白的成员。此外,MalK参与了两种不同的调控途径,调节了MalFGK的基因表达和运输活性。我们已经创建了一组malK突变来分析蛋白质的结构和折叠。这些突变分布在malK中,都是类似的31个密码子插入。每个突变体在麦芽糖转运和麦芽糖依赖性调节中的功能被表征。此外,我们利用灵敏的生化分析将MalK插入突变体分为两类:MalFGK复杂组装熟练和复杂组装缺陷。汇编精通类中包含插入的区域应与MalFGK复合体中表面暴露的MalK区域相对应。装配缺陷突变体的受影响区域可能涉及复合体内的关键结构接触。一个突变体在最终MalFGK复合物寡聚化之前的中间阶段明显阻断组装。这项工作有助于ABC转运蛋白的分析和异质低聚膜蛋白组装过程的研究。
MalK is a peripheral cytoplasmic membrane protein that has multiple activities in Escherichia coli. It associates with integral cytoplasmic membrane proteins MalF and MalG to form the maltose transport complex (MalFGK), a member of the ATP-binding cassette (ABC) superfamily of proteins. In addition, MalK participates in two different regulatory pathways which modulate mal gene expression and MalFGK transport activity. We have created a set of malK mutations for analysis of the protein's structure and folding. These mutations, distributed throughout malK, are all similar insertions of 31 codons. The ability of each mutant to function in maltose transport and MalK-dependent regulation was characterized. Furthermore, we have exploited a sensitive biochemical assay to classify our MalK insertion mutants into two additional categories: MalFGK complex assembly proficient and complex assembly defective. The regions containing the insertions in the assembly-proficient class should correspond to areas within MalK that are surface exposed within the MalFGK complex. Affected regions in assembly-deficient mutants may be involved in critical structural contacts within the complex. One mutant apparently blocks assembly at an intermediate stage prior to oligomerization of the final MalFGK complex. This work contributes to the analysis of ABC transport proteins and to the study of the assembly process for hetero-oligomeric membrane proteins.
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