The Net Charge of the First 18 Residues of the Mature Sequence Affects Protein Translocation across the Cytoplasmic Membrane of Gram-Negative Bacteria

Andrey V. Kajava; Sergey N. Zolov; Andrey E. Kalinin; Marina A. Nesmeyanova

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The Net Charge of the First 18 Residues of the Mature Sequence Affects Protein Translocation across the Cytoplasmic Membrane of Gram-Negative Bacteria

机译：成熟序列的前18个残基的净电荷影响跨革兰氏阴性细菌细胞质膜的蛋白质转运。

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This statistical study shows that in proteins of gram-negative bacteria exported by the Sec-dependent pathway, the first 14 to 18 residues of the mature sequences have the highest deviation between the observed and expected net charge distributions. Moreover, almost all sequences have either neutral or negative net charge in this region. This rule is restricted to gram-negative bacteria, since neither eukaryotic nor gram-positive bacterial exported proteins have this charge bias. Subsequent experiments performed with a series ofEscherichia coli alkaline phosphatase mutants confirmed that this charge bias is associated with protein translocation across the cytoplasmic membrane. Two consecutive basic residues inhibit translocation effectively when placed within the first 14 residues of the mature protein but not when placed in positions 19 and 20. The sensitivity to arginine partially reappeared again 30 residues away from the signal sequence. These data provide new insight into the mechanism of protein export in gram-negative bacteria and lead to practical recommendations for successful secretion of hybrid proteins.

机译：这项统计研究表明，在通过Sec依赖性途径输出的革兰氏阴性细菌的蛋白质中，成熟序列的前14至18个残基在观察到的和预期的净电荷分布之间具有最高的偏差。此外，几乎所有序列在该区域均具有中性或负性净电荷。该规则仅限于革兰氏阴性细菌，因为真核或革兰氏阳性细菌输出蛋白都没有这种电荷偏向。随后对一系列大肠埃希氏大肠杆菌碱性磷酸酶突变体进行的实验证实，这种电荷偏倚与蛋白质跨细胞质膜的转运有关。当放置在成熟蛋白的前14个残基中时，两个连续的碱性残基有效地抑制了易位，但是当放置在19和20位时则无效。对精氨酸的敏感性又部分地从信号序列中再次出现了30个残基。这些数据为革兰氏阴性细菌中蛋白质输出的机理提供了新的见识，并为成功分泌杂交蛋白提供了实用建议。

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《Journal of bacteriology》 |2000年第8期|共7页
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Andrey V. Kajava; Sergey N. Zolov; Andrey E. Kalinin; Marina A. Nesmeyanova;
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1. Protein translocation across the inner membrane of Gram-negative bacteria: The Sec and Tat dependent protein transport pathways [J] . KudvaR., DenksK., KuhnP., Research in Microbiology . 2013,第6期

机译：跨革兰氏阴性细菌内膜的蛋白质转运：Sec和Tat依赖性蛋白质转运途径
2. Altered localization of HrpZ in Pseudomonas syringae pv. syringae hrp mutants suggests that different components of the type III secretion pathway control protein translocation across the inner and outer membranes of gram-negative bacteria. [J] . A O Charkowski, H C Huang, A Collmer Journal of bacteriology . 1997,第12期

机译：丁香假单胞菌PV中HrpZ的本地化。丁香属植物hrp突变体表明，III型分泌途径的不同成分控制着蛋白质在革兰氏阴性细菌的内膜和外膜上的移位。
3. Prediction of lipid-binding regions in cytoplasmic and extracellular loops of membrane proteins as exemplified by protein translocation membrane proteins [J] . KellerR.C.A. The Journal of Membrane Biology: An International Journal for Studies on the Structure, Function & Genesis of Biomembranes . 2013,第1期

机译：预测膜蛋白胞质和细胞外环中的脂质结合区，以蛋白易位膜蛋白为例
4. Antibacterial performance of graphene oxide complemented with pluronic F-127 on physiologically mature gram-negative bacteria [C] . H. Enis Karahan, Christian Wiraja, N. Farahin Binte Nasrun, International Conference on Nanotechnology . 2017

机译：氧化石墨烯与普朗尼克F-127互补对生理成熟革兰氏阴性细菌的抗菌性能
5. The role of cytoplasmic chaperones in the biogenesis, maturation, and degradation of cytoplasmic and integral membrane proteins. [D] . Ahner, Annette. 2005

机译：细胞质伴侣在细胞质和整合膜蛋白的生物发生，成熟和降解中的作用。
6. The Net Charge of the First 18 Residues of the Mature Sequence Affects Protein Translocation across the Cytoplasmic Membrane of Gram-Negative Bacteria [O] . Andrey V. Kajava, Sergey N. Zolov, Andrey E. Kalinin, 2000

机译：成熟序列的前18个残基的净电荷影响跨革兰氏阴性菌细胞质膜的蛋白质转运。
7. The Net Charge of the First 18 Residues of the Mature Sequence Affects Protein Translocation across the Cytoplasmic Membrane of Gram-Negative Bacteria [O] . Kajava, Andrey V., Zolov, Sergey N., Kalinin, Andrey E., 2000

机译：成熟序列的前18个残基的净电荷影响跨革兰氏阴性细菌细胞质膜的蛋白质转运。

The Net Charge of the First 18 Residues of the Mature Sequence Affects Protein Translocation across the Cytoplasmic Membrane of Gram-Negative Bacteria

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