Theoretical and computational investigation of flagellin translocation and bacterial flagellum growth.

The bacterial flagellum is a self-assembling filament, which bacteria use for swimming. It is built from tens of thousands of flagellin monomers in a self-assembly process that involves translocation of the monomers through the flagellar interior, a channel, to the growing tip. Flagellum monomers are pumped into the filament at the base, move unfolded along the channel and then bind to the tip of the filament, thereby extending the growing flagellum. The flagellin translocation process, due to the flagellum maximum length of 20 μm, is an extreme example of protein transport through channels. Here, we derive a model for flagellin transport through the long confining channel, testing the key assumptions of the model through molecular dynamics simulations that also furnish system parameters needed for quantitative description. Together, mathematical model and molecular dynamics simulations explain why the growth rate of flagellar filaments decays exponentially with filament length and why flagellum growth ceases at a certain maximum length.

[1]  Peter Michaely,et al.  Nanospring behaviour of ankyrin repeats , 2006, Nature.

[2]  K. Namba Roles of partly unfolded conformations in macromolecular self‐assembly , 2001, Genes to cells : devoted to molecular & cellular mechanisms.

[3]  K. Namba,et al.  Structure of the core and central channel of bacterial flagella , 1989, Nature.

[4]  D. Case,et al.  Exploring protein native states and large‐scale conformational changes with a modified generalized born model , 2004, Proteins.

[5]  M. Ryan,et al.  Translocation of Proteins into Mitochondria , 2001, IUBMB life.

[6]  A. Kitao,et al.  Thermal Unfolding Simulations of Bacterial Flagellin: Insight into its Refolding Before Assembly , 2008, Biophysical journal.

[7]  G. Oster,et al.  What drives the translocation of proteins? , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Zaida Luthey-Schulten,et al.  MultiSeq: unifying sequence and structure data for evolutionary analysis , 2006, BMC Bioinformatics.

[9]  Takashi Kumasaka,et al.  Structure of the bacterial flagellar protofilament and implications for a switch for supercoiling , 2001, Nature.

[10]  Domain movements of HAP2 in the cap–filament complex formation and growth process of the bacterial flagellum , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Sung,et al.  Polymer Translocation through a Pore in a Membrane. , 1996, Physical review letters.

[12]  山川 裕巳,et al.  Modern theory of polymer solutions , 1971 .

[13]  K. Schulten,et al.  The Roles of Pore Ring and Plug in the SecY Protein-conducting Channel , 2008, The Journal of general physiology.

[14]  Randy Schekman,et al.  Protein Translocation Across Biological Membranes , 2005, Science.

[15]  G. McClelland,et al.  Atomic-scale friction of a tungsten tip on a graphite surface. , 1987, Physical review letters.

[16]  Shin-Ichi Aizawa,et al.  Type III secretion systems and bacterial flagella: Insights into their function from structural similarities , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Adler,et al.  Purification of Intact Flagella from Escherichia coli and Bacillus subtilis , 1971, Journal of bacteriology.

[18]  S. Lowen The Biophysical Journal , 1960, Nature.

[19]  K Namba,et al.  The structure of the R-type straight flagellar filament of Salmonella at 9 A resolution by electron cryomicroscopy. , 1995, Journal of molecular biology.

[20]  R. Macnab,et al.  Domain organization and function of Salmonella FliK, a flagellar hook-length control protein. , 2004, Journal of molecular biology.

[21]  T. Baumberger,et al.  Creeplike relaxation at the interface between rough solids under shear , 1996 .

[22]  川勝 年洋 Statistical physics of polymers : an introduction , 2004 .

[23]  S. Smith,et al.  Folding-unfolding transitions in single titin molecules characterized with laser tweezers. , 1997, Science.

[24]  S. Goodwin,et al.  Simulation studies of polymer translocation through a channel. , 2003, Physical review. E, Statistical, nonlinear, and soft matter physics.

[25]  A. Fasolino,et al.  Velocity dependence of atomic-scale friction: A comparative study of the one- and two-dimensional Tomlinson model , 2005 .

[26]  Koji Yonekura,et al.  Building the atomic model for the bacterial flagellar filament by electron cryomicroscopy and image analysis. , 2005, Structure.

[27]  D. DeRosier,et al.  Structure of Bacterial Flagellar Filaments at 11 Å Resolution: Packing of the α-Helices , 1995 .

[28]  M. Muthukumar,et al.  Simulation of polymer translocation through protein channels. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[29]  T. Iino Assembly of Salmonella flagellin in vitro and in vivo. , 1974, Journal of supramolecular structure.

[30]  S. Spragg Biophysical chemistry , 1979, Nature.

[31]  How static is static friction? , 2008, Proceedings of the National Academy of Sciences.

[32]  K Schulten,et al.  VMD: visual molecular dynamics. , 1996, Journal of molecular graphics.

[33]  Rob Phillips,et al.  Forces during bacteriophage DNA packaging and ejection. , 2004, Biophysical journal.

[34]  Laxmikant V. Kalé,et al.  Scalable molecular dynamics with NAMD , 2005, J. Comput. Chem..

[35]  Driven polymer translocation through a nanopore: A manifestation of anomalous diffusion , 2007, cond-mat/0702463.

[36]  D. Lubensky,et al.  Driven polymer translocation through a narrow pore. , 1999, Biophysical journal.

[37]  Klaus Schulten,et al.  Coarse-grained molecular dynamics simulations of a rotating bacterial flagellum. , 2006, Biophysical journal.

[38]  F. Heslot,et al.  Creep, stick-slip, and dry-friction dynamics: Experiments and a heuristic model. , 1994, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[39]  D. Blair Flagellar movement driven by proton translocation , 2003, FEBS letters.

[40]  Wolfgang A. Linke,et al.  Reverse engineering of the giant muscle protein titin , 2002, Nature.

[41]  Jens Michaelis,et al.  Mechanism of Force Generation of a Viral DNA Packaging Motor , 2005, Cell.

[42]  K. Schulten,et al.  Steered molecular dynamics and mechanical functions of proteins. , 2001, Current opinion in structural biology.

[43]  T. Lino Assembly of Salmonella flagellin in vitro and in vivo. , 1974 .

[44]  Flagellar elongation: an example of controlled growth. , 1974, Journal of theoretical biology.

[45]  E. Levy Flagellar elongation as a moving boundary problem. , 1974, Bulletin of mathematical biology.

[46]  J. C. Jaeger,et al.  Conduction of Heat in Solids , 1952 .

[47]  Alexander D. MacKerell,et al.  All-atom empirical potential for molecular modeling and dynamics studies of proteins. , 1998, The journal of physical chemistry. B.

[48]  R. Macnab,et al.  Substrate Specificity Classes and the Recognition Signal for Salmonella Type III Flagellar Export , 2003, Journal of bacteriology.

[49]  K. Namba,et al.  Complete atomic model of the bacterial flagellar filament by electron cryomicroscopy , 2003, Nature.

[50]  B. Dobberstein,et al.  Common Principles of Protein Translocation Across Membranes , 1996, Science.

[51]  W. L. Jorgensen,et al.  Comparison of simple potential functions for simulating liquid water , 1983 .