APh161 - Lecture 15: Molecular MotorsSlide 2Rogues Gallery of Motor Action: Rotary MotorsRogues Gallery of Motor Action: TranslocationRogues Gallery of Motor Action: Translational Motor 1Rogues Gallery of Motor Action: Translational Motor 2 - MusclesSlide 7Organellar TransportRogues Gallery of Motor Action: Translational Motor 2Slide 10How We Know: Gliding Motility AssaysDynamics of Molecular MotorsStepping KineticsKinesin DataKinesin Data continuedKinesin Randomness DataMotors and StatesWhat do we mean by the states?Translocation MotorsMeasurements on Rate of DNA EjectionSlide 21Slide 22Slide 23APh161 - Lecture 15: Molecular APh161 - Lecture 15: Molecular MotorsMotorsRob PhillipsCalifornia Institute of TechnologyMitotic Spindle OrganizationCilia and Flagella Assembly and DynamicsFormation of Golgi and ER NetworksVesicle TransportMolecular MotorsMolecular MotorsRogues Gallery of Motor Action: Rotary Rogues Gallery of Motor Action: Rotary MotorsMotorsShow Berg movieShow Yasuda et al. movieRogues Gallery of Motor Action: Rogues Gallery of Motor Action: TranslocationTranslocationRogues Gallery of Motor Action: Rogues Gallery of Motor Action: Translational Motor 1Translational Motor 1Rogues Gallery of Motor Action: Rogues Gallery of Motor Action: Translational Motor 2 - MusclesTranslational Motor 2 - MusclesSee Hugh Huxley review on websiteRogues Gallery of Motor Action: Rogues Gallery of Motor Action: Translational Motor 2 - MusclesTranslational Motor 2 - MusclesHeuser lab – Washington UniversityOrganellar Transport Organellar Transport (Hirokawa, Science 1998)Rogues Gallery of Motor Action: Rogues Gallery of Motor Action: Translational Motor 2Translational Motor 2Myosin VSpeed: 350 nm/s ATPase: 5.0 1/sKinesinSpeed: 850 nm/s ATPase: 44.0 1/sDyneinSpeed: -1250 nm/s ATPase: 2.0 1/s**ALL INVOLVED IN VESICLE TRANSPORT**Translational MotorsTranslational MotorsHow We Know: Gliding Motility AssaysHow We Know: Gliding Motility AssaysShow Vale movieDynamics of Molecular MotorsDynamics of Molecular MotorsScience, Vol 300, Issue 5628, 2061-2065, 27 June 2003 - Yildiz et al.Show Gelles, Selvin movieStepping KineticsStepping KineticsSpudich et al., PNAS 2000Kinesin DataKinesin DataFig. 3. Kinesin velocity as a function of [ATP] under external loads, F, fixed by a force clamp. The plots, from the top down, are for F = 0, 1.05, 3.59, 4.60, and 5.63 pN, respectively. Data from Block and colleagues (9): solid curves, N = 2 fits; dashed curves, N = 2 predictions (see text) (Fisher and Kolomeisky, PNAS)Kinesin Data continuedKinesin Data continuedFig. 4. Fits to the data of Block and colleagues (9) (and predictions) for velocity as a function of load for fixed concentrations of ATP. Note the inflection points at low [ATP] and convex profile at saturating [ATP]. (Fisher and Kolomeisky, PNAS)Kinesin Randomness DataKinesin Randomness DataFig. 5. Randomness data from Block and colleagues (9) and theoretical fits (A) as a function of external load, F, at fixed [ATP] (note that the two data points at F 5.7 pN and [ATP] = 2 mM appear separately in Block and coworkers: see figure 4 a and b of ref. 9, respectively) and (B) as a function of [ATP] at fixed loads of, from top down, F = 5.69 pN ( ), 5.35 and 4.60 pN (dashed-line predictions), 3.59 pN ( ), and 1.05 pN ( ).Motors and StatesMotors and StatesWhat do we mean by the states?What do we mean by the states?Translocation MotorsTranslocation MotorsMeasurements on Rate of DNA EjectionMeasurements on Rate of DNA EjectionDNA injection from T5 into vesicleInjection rate in λ (Baldeschweiler et al.) (Bohm et al.)Measurements on Rate of DNA EjectionMeasurements on Rate of DNA EjectionPhage Hypothesized MechanismGenome Length (kbp)Ejection time (sec)Av. Ejection rate (kbp/sec)lambda Pressure 48.5 60 0.8T4 Pressure 169 30 5.6T7 Enzyme 40 600 0.06T5 Pressure+Enzyme121 360 0.3phi29 Pressure+ Enzyme 19 1800 0.05Calculating the Ejection Time Calculating the Ejection Time Calculating the Ejection Time Calculating the Ejection Time The mean first passage time isThe Time of EjectionThe Time of EjectionreceptorvesicleNo protein
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