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    Article: Switching Statistics of a Flagellar Motor: First-Passage Time and Dynamic Binding
    Qi Wen, Guanglai Li, Jay X. Tang, Greg Huber
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    ABSTRACT: Bacterial chemotaxis relies on the clockwise-to-counterclockwise switching (and the reverse CCW-to-CW switching) of the flagellar motor’s rotational direction. Yet, despite its central role, the switch mechanism remains poorly understood. Here we study the switch statistics of the flagellar motor using computer simulations that combine first-passage time ideas and the property of dynamic binding of CheY-P to FliM. The simulated clockwise bias and switching frequency as a function of CheY-P concentration agree with the experimental results for E. coli. Moreover, the simulation results show a robust peak, which is present in some experimental reports, in the distribution of the time intervals of both clockwise and counterclockwise rotation.
    Journal of Statistical Physics 04/2012; 128(1):257-267. · 1.40 Impact Factor
  • Article: Surface contact stimulates the just-in-time deployment of bacterial adhesins.
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    ABSTRACT: The attachment of bacteria to surfaces provides advantages such as increasing nutrient access and resistance to environmental stress. Attachment begins with a reversible phase, often mediated by surface structures such as flagella and pili, followed by a transition to irreversible attachment, typically mediated by polysaccharides. Here we show that the interplay between pili and flagellum rotation stimulates the rapid transition between reversible and polysaccharide-mediated irreversible attachment. We found that reversible attachment of Caulobacter crescentus cells is mediated by motile cells bearing pili and that their contact with a surface results in the rapid pili-dependent arrest of flagellum rotation and concurrent stimulation of polar holdfast adhesive polysaccharide. Similar stimulation of polar adhesin production by surface contact occurs in Asticcacaulis biprosthecum and Agrobacterium tumefaciens. Therefore, single bacterial cells respond to their initial contact with surfaces by triggering just-in-time adhesin production. This mechanism restricts stable attachment to intimate surface interactions, thereby maximizing surface attachment, discouraging non-productive self-adherence, and preventing curing of the adhesive.
    Molecular Microbiology 11/2011; 83(1):41-51. · 5.01 Impact Factor
  • Article: Accumulation of swimming bacteria near a solid surface.
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    ABSTRACT: We measured the distribution of a forward swimming strain of Caulobacter crescentus near a surface using a three-dimensional tracking technique based on dark field microscopy and found that the swimming bacteria accumulate heavily within a micrometer from the surface. We attribute this accumulation to frequent collisions of the swimming cells with the surface, causing them to align parallel to the surface as they continually move forward. The extent of accumulation at the steady state is accounted for by balancing alignment caused by these collisions with the rotational Brownian motion of the micrometer-sized bacteria. We performed a simulation based on this model, which reproduced the measured results. Additional simulations demonstrate the dependence of accumulation on swimming speed and cell size, showing that longer and faster cells accumulate more near a surface than shorter and slower ones do.
    Physical Review E 10/2011; 84(4 Pt 1):041932. · 2.26 Impact Factor
  • Article: Surface adsorption and hopping cause probe-size-dependent microrheology of actin networks.
    Jun He, Jay X Tang
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    ABSTRACT: A network of filaments formed primarily by the abundant cytoskeletal protein actin gives animal cells their shape and elasticity. The rheological properties of reconstituted actin networks have been studied by tracking micron-sized probe beads embedded within the networks. We investigate how microrheology depends on surface properties of probe particles by varying the stickiness of their surface. For this purpose, we chose carboxylate polystyrene (PS) beads, silica beads, bovine serum albumin (BSA) -coated PS beads, and polyethylene glycol (PEG) -grafted PS beads, which show descending stickiness to actin filaments, characterized by confocal imaging and microrheology. Probe size dependence of microrheology is observed for all four types of beads. For the slippery PEG beads, particle-tracking microrheology detects weaker networks using smaller beads, which tend to diffuse through the network by hopping from one confinement "cage" to another. This trend is reversed for the other three types of beads, for which microrheology measures stiffer networks for smaller beads due to physisorption of nearby filaments to the bead surface. We explain the probe size dependence with two simple models. We also evaluate depletion effect near nonadsorption bead surface using quantitative image analysis and discuss the possible impact of depletion on microrheology. Analysis of these effects is necessary in order to accurately define the actin network rheology both in vitro and in vivo.
    Physical Review E 04/2011; 83(4 Pt 1):041902. · 2.26 Impact Factor
  • Article: Nitric oxide reduces sickle hemoglobin polymerization: potential role of nitric oxide-induced charge alteration in depolymerization.
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    ABSTRACT: We previously demonstrated that inhaling nitric oxide (NO) increases the oxygen affinity of sickle red blood cells (RBCs) in patients with sickle cell disease (SCD). Our recent studies found that NO lowered the P(50) values of sickle hemoglobin (HbS) hemolysates but did not increase methemoglobin (metHb) levels, supporting the role of NO, but not metHb, in the oxygen affinity of HbS. Here we examine the mechanism by which NO increases HbS oxygen affinity. Because anti-sickling agents increase sickle RBC oxygen affinity, we first determined whether NO exhibits anti-sickling properties. The viscosity of HbS hemolysates, measured by falling ball assays, increased upon deoxygenation; NO treatment reduced the increment. Multiphoton microscopic analyses showed smaller HbS polymers in deoxygenated sickle RBCs and HbS hemolysates exposed to NO. These results suggest that NO inhibits HbS polymer formation and has anti-sickling properties. Furthermore, we found that HbS treated with NO exhibits an isoelectric point similar to that of HbA, suggesting that NO alters the electric charge of HbS. NO-HbS adducts had the same elution time as HbA upon high performance liquid chromatography analysis. This study demonstrates that NO may disrupt HbS polymers by abolishing the excess positive charge of HbS, resulting in increased oxygen affinity.
    Archives of Biochemistry and Biophysics 03/2011; 510(1):53-61. · 2.93 Impact Factor

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