Self-assembly and recrystallization of bacterial S-layer proteins at silicon supports imaged in real time by atomic force microscopy
Self-assembly and recrystallization of bacterial S-layer proteins at silicon supports imaged in real time by atomic force microscopy
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DOI：
10.1111/j.1365-2818.2003.01270.x
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发表时间：
2003-12-01
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影响因子：
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通讯作者：
Sleytr, UB
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文献类型：
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作者：
Györvary, ES;Stein, O;Sleytr, UB
利用原子力显微镜（AFM）对球形芽孢杆菌CCM 2177的SbpA蛋白在硅载体（疏水性、非等离子体处理和亲水性、O-2等离子体处理的硅载体）上的自组装进行了真实的实时成像。一个封闭的马赛克层组成的小晶体（直径小于200纳米）形成在疏水性硅支持，而一个连贯的晶格组成的大域（2-10 μ m的大小）产生在O-2等离子体处理，亲水性硅晶片。所形成的层的结构是在疏水性硅处的单层（高度为9 nm）和在亲水性硅处的双层（高度为15 nm）。原位原子力显微镜测量证实了离子键在硅支持的结晶SbpA层的形成中的重要性。断裂的蛋白质亚基与金属螯合剂从晶格的SbpA原位可视化的AFM。用氨基-氨基或氨基-羧基定向交联剂交联SbpA层可以提高固体支撑SbpA层的稳定性。
The self-assembly of bacterial surface-layer (S-layer) proteins (SbpA of Bacillus sphaericus CCM 2177) at silicon supports (hydrophobic, non-plasma-treated and hydrophilic, O-2 plasma-treated silicon supports) was imaged in real time by atomic force microscopy (AFM). A closed mosaic layer consisting of small crystals (less than 200 nm in diameter) was formed at a hydrophobic silicon support, whereas a coherent crystalline lattice consisting of large domains (2-10 mum in size) was generated at O-2 plasma-treated, hydrophilic silicon wafers. The structure of the formed layers was a monolayer (9 nm in height) at the hydrophobic silicon and a bilayer (15 nm in height) at the hydrophilic silicon. In situ AFM measurements confirmed the importance of ionic bonds in the formation of crystalline SbpA layers at silicon supports. Rupture of the protein subunits with a metal chelator from the crystalline lattice of SbpA was visualized in situ by AFM. The stability of solid-supported SbpA layers could be enhanced by cross-linking the S-layers with amino-amino or amino-carboxyl group directed cross-linkers.