Structure-Function Analysis of the Two-Peptide Bacteriocin Plantaricin EF.
Structure-Function Analysis of the Two-Peptide Bacteriocin Plantaricin EF.
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DOI：
10.1021/acs.biochem.6b00588
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发表时间：
2016-09-13
期刊：
影响因子：
2.9
通讯作者：
Kristiansen, Per Eugen
中科院分区：
文献类型：
--
作者：
Ekblad, Bie;Kyriakou, Panagiota K.;Oppegard, Camilla;Nissen-Meyer, Jon;Kaznessis, Yiannis N.;Kristiansen, Per Eugen
Plantaricin EF是一种双肽细菌素，依赖于两个不同的多肽(PlnE和PlnF)的互补作用而发挥作用。单个多肽的结构以前已经通过核磁共振波谱进行了分析(Fimland，N.et al.()，，−)，但细菌素的结构和这两个肽如何相互作用尚未确定。到目前为止，所有已鉴定的双肽细菌素都含有GxxxG基序。这些基序与GxxxG类基序一起，在膜蛋白中介导螺旋-螺旋相互作用。我们对PlnE和PlnF中的所有GxxxG和GxxxG样基序进行了突变，以确定这些基序中是否有任何基序对抗菌活性重要，从而可能对PlnE和PlnF之间的相互作用起重要作用。此外，还对PlnE和PlnF中的芳香族氨基酸Tyr和Trp进行了取代，并构建了4个融合多肽，以研究PlnE和PlnF在靶细胞膜中的相对定位。结果表明，PlnE和PlnF以反平行的方式相互作用，PlnE的C端和PlnF的N端位于靶细胞膜的外侧，PlnE的N端和PlnF的C端位于靶细胞膜的内侧。PlnE中6位的芳香族残基的偏好表明该残基定位在内部细胞上的膜界面内或附近。GxxxG基序的突变表明PlnE中的G5xxxG9基序和PlnF中的S26xxxG30基序参与了螺旋-螺旋相互作用。原子分子动力学模拟的结构模型与实验结果一致，证实了膜中结构及其取向的稳定性。模拟证实了预期的相互作用，并揭示了进一步提高拟议结构稳定性的额外相互作用。
Plantaricin EF is a two-peptide bacteriocin that depends on the complementary action of two different peptides (PlnE and PlnF) to function. The structures of the individual peptides have previously been analyzed by nuclear magnetic resonance spectroscopy (Fimland, N. et al. (), , −), but the bacteriocin structure and how the two peptides interact have not been determined. All two-peptide bacteriocins identified so far contain GxxxG motifs. These motifs, together with GxxxG-like motifs, are known to mediate helix–helix interactions in membrane proteins. We have mutated all GxxxG and GxxxG-like motifs in PlnE and PlnF in order to determine if any of these motifs are important for antimicrobial activity and thus possibly for interactions between PlnE and PlnF. Moreover, the aromatic amino acids Tyr and Trp in PlnE and PlnF were substituted, and four fusion polypeptides were constructed in order to investigate the relative orientation of PlnE and PlnF in target cell membranes. The results obtained with the fusion polypeptides indicate that PlnE and PlnF interact in an antiparallel manner and that the C-terminus of PlnE and N-terminus of PlnF are on the outer part of target cell membranes and the N-terminus of PlnE and C-terminus of PlnF are on the inner part. The preference for an aromatic residue at position 6 in PlnE suggests a positioning of this residue in or near the membrane interface on the cells inside. Mutations in the GxxxG motifs indicate that the G5xxxG9 motif in PlnE and the S26xxxG30 motif in PlnF are involved in helix–helix interactions. Atomistic molecular dynamics simulation of a structural model consistent with the results confirmed the stability of the structure and its orientation in membranes. The simulation approved the anticipated interactions and revealed additional interactions that further increase the stability of the proposed structure.
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影响因子：
3.2
作者：
AXELSSON, L;HOLCK, A
通讯作者：
HOLCK, A
影响因子：
3.3
作者：
Klauda, Jeffery B.;Venable, Richard M.;Freites, J. Alfredo;O'Connor, Joseph W.;Tobias, Douglas J.;Mondragon-Ramirez, Carlos;Vorobyov, Igor;MacKerell, Alexander D., Jr.;Pastor, Richard W.
通讯作者：
Pastor, Richard W.
DOI：
10.1002/jbmte.390020409
发表时间：
1960-01-01
期刊：
JOURNAL OF BIOCHEMICAL AND MICROBIOLOGICAL TECHNOLOGY AND ENGINEERING
影响因子：
--
作者：
EDEBO, L
通讯作者：
EDEBO, L
影响因子：
3.2
作者：
Moll, GN;van den Akker, E;Driessen, AJM
通讯作者：
Driessen, AJM
影响因子：
3.6
作者：
Kjos, Morten;Oppegard, Camilla;Kristensen, Tom
通讯作者：
Kristensen, Tom