Variations in the nanomechanical properties of virulent and avirulent Listeria monocytogenes

Abstract

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 a high density polymer surface 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 that the 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.