Correlative Super-Resolution Optical and Atomic Force Microscopy Reveals Relationships Between Bacterial Cell Wall Architecture and Synthesis in Bacillus subtilis.
Correlative Super-Resolution Optical and Atomic Force Microscopy Reveals Relationships Between Bacterial Cell Wall Architecture and Synthesis in Bacillus subtilis.
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DOI：
10.1021/acsnano.1c04375
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发表时间：
2021-10-26
期刊：
影响因子：
17.1
通讯作者：
Hobbs JK
中科院分区：
文献类型：
--
作者：
Tank RKG;Lund VA;Kumar S;Turner RD;Lafage L;Pasquina Lemonche L;Bullough PA;Cadby A;Foster SJ;Hobbs JK
了解细菌如何生长和分裂需要深入了解超微结构的分子水平动力学和组成成分的化学。原子力显微镜（AFM）可以提供生物系统的近分子分辨率图像，但通常提供有限的化学信息。相反，虽然超分辨率光学显微镜可以定位特定的分子和化学物质，但很难获得分子背景的信息。在这里，我们联合收割机这些方法结合到STORMForce（随机光学重建与原子力显微镜）和补充SIMForce（结构照明与原子力显微镜），映射细菌细胞壁结构大分子，肽聚糖的合成，在杆状细菌枯草芽孢杆菌的生长和分裂过程中。使用“可点击”的D-氨基酸掺入，我们荧光标记和空间定位的肽聚糖合成的一个短的和受控的时期，并将此信息与高分辨率AFM的所得架构。在分裂过程中，隔合成发生在其发展的表面，这表明一个两阶段的过程与合并的前缘和相当大的填充后面。在生长过程中，杆的伸长通过合成带发生，间隔约300 nm，并且对应于由AFM揭示的内部细胞壁的致密区域。结合超分辨率光学和原子力显微镜可以提供深入了解细菌结构生物聚合物的复杂结构的合成过程。
Understanding how bacteria grow and divide requires insight into both the molecular-level dynamics of ultrastructure and the chemistry of the constituent components. Atomic force microscopy (AFM) can provide near molecular resolution images of biological systems but typically provides limited chemical information. Conversely, while super-resolution optical microscopy allows localization of particular molecules and chemistries, information on the molecular context is difficult to obtain. Here, we combine these approaches into STORMForce (stochastic optical reconstruction with atomic force microscopy) and the complementary SIMForce (structured illumination with atomic force microscopy), to map the synthesis of the bacterial cell wall structural macromolecule, peptidoglycan, during growth and division in the rod-shaped bacterium Bacillus subtilis. Using “clickable” d-amino acid incorporation, we fluorescently label and spatially localize a short and controlled period of peptidoglycan synthesis and correlate this information with high-resolution AFM of the resulting architecture. During division, septal synthesis occurs across its developing surface, suggesting a two-stage process with incorporation at the leading edge and with considerable in-filling behind. During growth, the elongation of the rod occurs through bands of synthesis, spaced by ∼300 nm, and corresponds to denser regions of the internal cell wall as revealed by AFM. Combining super-resolution optics and AFM can provide insights into the synthesis processes that produce the complex architectures of bacterial structural biopolymers.
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DOI：
10.1002/anie.201206749
发表时间：
2012-12-07
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Kuru E;Hughes HV;Brown PJ;Hall E;Tekkam S;Cava F;de Pedro MA;Brun YV;VanNieuwenhze MS
通讯作者：
VanNieuwenhze MS
影响因子：
10.8
作者：
Odermatt, Pascal D.;Shivanandan, Arun;Fantner, Georg E.
通讯作者：
Fantner, Georg E.
影响因子：
5.2
作者：
Dersch, Simon;Mehl, Johanna;Graumann, Peter L.
通讯作者：
Graumann, Peter L.
影响因子：
16.6
作者：
Radkov AD;Hsu YP;Booher G;VanNieuwenhze MS
通讯作者：
VanNieuwenhze MS
影响因子：
28.3
作者：
Dion, Michael F.;Kapoor, Mrinal;Garner, Ethan C.
通讯作者：
Garner, Ethan C.