Mapping the Sites of Interaction between SecY and SecE by Cysteine Scanning Mutagenesis*

Mapping the Sites of Interaction between SecY and SecE by Cysteine Scanning Mutagenesis*
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通过半胱氨酸扫描诱变绘制 SecY 和 SecE 之间的相互作用位点*

DOI:
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发表时间:
2001
影响因子:
4.8
通讯作者:
A. Driessen
A. Driessen
中科院分区:
生物学2区
文献类型:
--
作者:
A. Veenendaal;C. van der Does;A. Driessen

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在大肠杆菌中,SecYEG复合物介导蛋白质的易位和膜整合。遗传和生化数据都表明SecY的几个跨膜片段(tms)与SecE相互作用。通过半胱氨酸扫描诱变,我们确定了SecY的TMS7和SecE的TMS3的分子间接触位点。SecY与SecE的交联表明,这些亚基在SecYEG复合物内以一对一的化学计量存在。SecE的TMS3中参与SecE二聚化的位点局限于特定的α-螺旋界面,并出现在低聚SecYEG复合物中。虽然交联可逆地使易位失活,但在将前蛋白插入易位通道后,SecY的TMS7和SecE的TMS3之间的接触保持不变。这些数据支持一个低聚易位通道模型,在该通道中,SecYEG复合物对通过SecE相互接触。
In Escherichia coli, the SecYEG complex mediates the translocation and membrane integration of proteins. Both genetic and biochemical data indicate interactions of several transmembrane segments (TMSs) of SecY with SecE. By means of cysteine scanning mutagenesis, we have identified intermolecular sites of contact between TMS7 of SecY and TMS3 of SecE. The cross-linking of SecY to SecE demonstrates that these subunits are present in a one-to-one stoichiometry within the SecYEG complex. Sites in TMS3 of SecE involved in SecE dimerization are confined to a specific α-helical interface and occur in an oligomeric SecYEG complex. Although cross-linking reversibly inactivates translocation, the contact between TMS7 of SecY and TMS3 of SecE remains unaltered upon insertion of the preprotein into the translocation channel. These data support a model for an oligomeric translocation channel in which pairs of SecYEG complexes contact each other via SecE.
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