Uncoiling mechanism of Klebsiella pneumoniae type 3 pili measured by using optical tweezers

Pili are bacterial appendages that play many important roles in bacterial behaviors, physiology and interaction with hosts. Via pili, bacteria are able to adhere to, migrate onto, and colonize on host cells, mechanically. Different from the most studied type 1 and P type pili, which are rigid and thick with an average of 6~7 nm in diameter, type 3 pili are relatively tiny (3-5 nm in diameter) and flexible, and their biophysical properties remains unclear. By using optical tweezers, we found that the elongation processes of type 3 pili are divided into three phases: (1) elastic elongation, (2) uncoiling elongation, and (3) intrinsic elongation, separately. Besides, the uncoiling force of the recombinant pili displayed on the surface of E. coli [pmrkABCDV1F] is measured 20 pN in average stronger than that of E. coli [pmrkABCDV1]. This suggests that pilin MrkF is involved in determining the mechanical properties of the type 3 pili.

[1]  N. Cafferini,et al.  Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. , 2003, Research in microbiology.

[2]  W. Greenleaf,et al.  High-resolution, single-molecule measurements of biomolecular motion. , 2007, Annual review of biophysics and biomolecular structure.

[3]  G Waksman,et al.  Chaperone-assisted pilus assembly and bacterial attachment. , 2000, Current opinion in structural biology.

[4]  S. Chu,et al.  Observation of a single-beam gradient force optical trap for dielectric particles. , 1986, Optics letters.

[5]  S. Clegg,et al.  Binding of the type 3 fimbriae of Klebsiella pneumoniae to human endothelial and urinary bladder cells , 1997, Infection and immunity.

[6]  Xiaohong Huang,et al.  Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells. , 2007, Micron.

[7]  S. Clegg,et al.  Nucleotide sequence and functions of mrk determinants necessary for expression of type 3 fimbriae in Klebsiella pneumoniae , 1991, Journal of bacteriology.

[8]  Hwei-Ling Peng,et al.  Characterization of the type 3 fimbriae with different MrkD adhesins: possible role of the MrkD containing an RGD motif. , 2006, Biochemical and biophysical research communications.

[9]  Andres F Oberhauser,et al.  The mechanical properties of E. coli type 1 pili measured by atomic force microscopy techniques. , 2006, Biophysical journal.

[10]  Esther Bullitt,et al.  Structure and assembly of P-pili: a protruding hinge region used for assembly of a bacterial adhesion filament. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[11]  H. Flyvbjerg,et al.  Power spectrum analysis for optical tweezers , 2004 .

[12]  Lee Makowski,et al.  Structural polymorphism of bacterial adhesion pili , 1995, Nature.

[13]  Michael P. Sheetz,et al.  Single pilus motor forces exceed 100 pN , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. W. Wright,et al.  Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. , 1986, Analytical biochemistry.

[15]  Michael P. Sheetz,et al.  Pilus retraction powers bacterial twitching motility , 2000, Nature.

[16]  Staffan Schedin,et al.  Physical properties of Escherichia coli P pili measured by optical tweezers. , 2004, Biophysical journal.

[17]  Olga Yakovenko,et al.  Uncoiling Mechanics of Escherichia coli Type I Fimbriae Are Optimized for Catch Bonds , 2006, PLoS biology.

[18]  Staffan Schedin,et al.  The unfolding of the P pili quaternary structure by stretching is reversible, not plastic , 2005, EMBO reports.

[19]  A. Ashkin Forces of a single-beam gradient laser trap on a dielectric sphere in the ray optics regime. , 1992, Methods in cell biology.

[20]  Viola Vogel,et al.  Shear‐dependent ‘stick‐and‐roll’ adhesion of type 1 fimbriated Escherichia coli , 2004, Molecular microbiology.