Theses and Dissertations from UMD
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New submissions to the thesis/dissertation collections are added automatically as they are received from the Graduate School. Currently, the Graduate School deposits all theses and dissertations from a given semester after the official graduation date. This means that there may be up to a 4 month delay in the appearance of a give thesis/dissertation in DRUM
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Item Theoretical Studies of the Workings of Processive Molecular Motors(2017) Vu, Huong Thuy; Thirumalai, Devarajan; Biophysics (BIPH); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Processive molecular motors, such as kinesins, myosins and helicases, take multiple discrete steps on linear polar tracks such as microtubules, filamentous actin, and DNA/RNA substrates. Insights into the mechanisms and functions of this important class of biological motors have been obtained through observations from single-molecule experiments and structural studies. Such information includes the distribution of n, the number of steps motors take before dissociating, and v, the motor velocity, in the presence and absence of an external resistive force from single molecule experiments; and different structures of different states of motors at different conditions. Based on those available data, this thesis focuses on using both analytical and computational theoretical tools to investigate the workings of processive motors. Two examples of processive motors considered here are kinesins that walk on microtubules while transporting vesicles, and helicases which translocate on DNA/RNA substrate while unwinding the helix substrate. New physical principles and predictions related to their motility emerge from the proposed theories. The most significant results reported in this thesis are: Exact and approximate equations for velocity distribution, P(v), and runlength distribution, P(n), have been derived. Application of the theory to kinesins shows that P(v) is non-Gaussian and bimodal at high resistive forces. This unexpected behavior is a consequence of the discrete spacing between the alpha/beta tubulins, the building blocks of microtubule. In the case of helicases, we demonstrate that P(v) of typical helicases T7 and T4 shows signatures of heterogeneity, inferred from large variations in the velocity from molecule to molecule. The theory is used to propose experiments in order to distinguish between different physical basis for heterogeneity. We generated a one-microsecond atomic simulation trajectory capturing the docking process of the neck-linker, a crucial element deemed to be important in the motility of Kinesin-1. The conformational change in the neck linker is important in the force generation in this type of motor. The simulations revealed new conformations of the neck-linker that have not been noted in previous structural studies of Kinesin-1, but which are demonstrated to be relevant to another superfamily member, Kinesin-5. By comparing the simulation results with currently available data, we suggest that the two superfamilies might actually share more similarities in the neck-linker docking process than previously thought.Item Molecular basis of the kinematics of the kinesin step(2012) Zhang, Zhechun; Thirumalai, Devarajan; Biophysics (BIPH); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Kinesin is an ATP-dependent cellular transporter that ferries cargos towards the plus-end of a microtubule. Despite significant advances in experiments, which have provided deep insights into the motility of kinesin, the molecular events that occur in a single step have not been fully resolved. In order to provide these details, this thesis develops a structure of the complex between kinesin and microtubule, and devises new simulation methods to probe the stepping kinetics over a wide range of conditions. Hundreds of molecular movies of kinesin walking on the microtubule are generated using coarse-grained simulation methods. Analysis of these movies shows that there are three major stages in the stepping kinetics of kinesin. In addition, an allosteric network within kinesin, responsible for controlling nucleotide release, is identified using microsecond all-atom simulations. These simulations are used to answer two important questions. First, does kinesin move by a "power stroke" or by diffusion? During a single step, the trailing head of the kinesin detaches from the microtubule, passes the microtubule-bound leading head, and attaches to the target binding site 16 nm away. The target binding site, however, is one of eight accessible binding sites on the microtubule. Is it possible that the "power stroke" (a large conformational change) in the leading head, pulls the trailing head into the neighborhood of the target binding site? This remained unclear because the fraction of the 16 nm step associated with the power stroke and diffusion had never been quantified. Second, how does the microtubule accelerate ADP release from kinesin, which is a key step in completing a single step? The ADP binding site of kinesin is more than 1.5 nm away from the microtubule binding surface. Therefore, the microtubule must affect the ADP binding site through an allosteric mechanism. However, the structural basis for transmitting signals through the underlying allosteric network was previously unknown. Analysis of hundreds of kinesin steps generated using coarse-grained simulations showed that the power stroke associated with the docking of the neck linker to the leading head, is responsible for only 4 nm of the 16 nm step, and the remaining 12 nm is covered by diffusion. However, the power stroke in the leading head constrains the diffusion of the trailing head, decreases the probability of side steps, and therefore biases the trailing head, to the target binding site. Additional all-atom simulations of the ADP-kinesin-microtubule complex, revealed a surprisingly simple allosteric network within kinesin that explains the acceleration of ADP release upon microtubule binding. The allosteric network also explains two additional experimental observations on ADP release from kinesin.