Park Bong-Jae, Abu-Lail Nehal I
Gene and Linda Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA 99164-2710, United States.
Soft Matter. 2010 Jan 1;6(16):3898-3909. doi: 10.1039/b927260g.
Atomic force microscopy (AFM) was used to quantify both the nanomechanical properties of pathogenic (ATCC 51776 & EGDe) and non-pathogenic (ATCC 15313 & HCC25) Listeria monocytogenes strains and the conformational properties of their surface biopolymers. The nanomechanical properties of the various L. monocytogenes strains were quantified in terms of Young's moduli of cells. To estimate Young's moduli, the classic Hertz model of contact mechanics and a modified version of it that takes into account substrate effects were used to fit the AFM nanoindentation-force measurements collected while pushing onto the bacterial surface biopolymer brush. When compared, the classic Hertz model always predicted higher Young's moduli values of bacterial cell elasticity compared to the modified Hertz model. On average, the modified Hertz model showed that virulent strains are approximately twice as rigid (88.1 ± 14.5 KPa) as the avirulent strains (47.3 ± 7.6 kPa). To quantify the conformational properties of L. monocytogenes' strains surface biopolymers, two models were used. First, the entropic-based, statistical mechanical, random walk formulation, the wormlike chain (WLC) model was used to estimate the elastic properties of the bacterial surface molecules. The WLC model results indicated that the virulent strains are characterized by a more flexible surface biopolymers as indicated by shorter persistence lengths (L(p) = 0.21 ± 0.08 nm) compared to the avirulent strains (L(p) = 0.24 ± 0.14 nm). Second, a steric model developed to describe the repulsive forces measured between the AFM tip and bacterial surface biopolymers indicated that the virulent strains are characterized by crowded and longer biopolymer brushes compared to those of the avirulent strains. Finally, scaling relationships developed for grafted polyelectrolyte brushes indicated L. monocytogenes strains' biopolymer brushes are charged. Collectively, our data indicate that the conformational properties of the bacterial surface biopolymers and their surface densities play an important role in controlling the overall bacterial cell elasticity.
原子力显微镜(AFM)用于量化致病性(ATCC 51776和EGDe)和非致病性(ATCC 15313和HCC25)单核细胞增生李斯特氏菌菌株的纳米力学性质及其表面生物聚合物的构象性质。各种单核细胞增生李斯特氏菌菌株的纳米力学性质通过细胞的杨氏模量来量化。为了估计杨氏模量,使用经典的赫兹接触力学模型及其考虑底物效应的修改版本来拟合在压向细菌表面生物聚合物刷时收集的AFM纳米压痕力测量值。比较时,经典赫兹模型预测的细菌细胞弹性杨氏模量值总是高于修改后的赫兹模型。平均而言,修改后的赫兹模型表明,有毒菌株的刚性(88.1±14.5 KPa)约为无毒菌株(47.3±7.6 kPa)的两倍。为了量化单核细胞增生李斯特氏菌菌株表面生物聚合物的构象性质,使用了两个模型。首先,基于熵的统计力学随机游走公式,即蠕虫状链(WLC)模型,用于估计细菌表面分子的弹性性质。WLC模型结果表明,与无毒菌株(L(p)=0.24±0.14 nm)相比,有毒菌株的表面生物聚合物更灵活,其持久长度更短(L(p)=0.21±0.08 nm)。其次,一个用于描述AFM尖端与细菌表面生物聚合物之间测量的排斥力的空间位阻模型表明,与无毒菌株相比,有毒菌株的生物聚合物刷更拥挤且更长。最后,为接枝聚电解质刷建立的标度关系表明单核细胞增生李斯特氏菌菌株的生物聚合物刷带电荷。总体而言,我们的数据表明,细菌表面生物聚合物的构象性质及其表面密度在控制细菌细胞的整体弹性方面起着重要作用。