Collective Dynamics of Kinesin-1 Motor Proteins Transporting a Common Load

Author(s):  
Adam G. Hendricks ◽  
Bogdan I. Epureanu ◽  
Edgar Meyho¨fer

Kinesin-1 is a motor protein essential to intracellular transport that converts the energy from ATP hydrolysis to directed movement along microtubules. Experimental and theoretical characterization of kinesin-1 has focused on single-molecule experiments. These experiments show that one motor is capable of transporting a cargo at speeds of about 1 μm/sec and maintaining contact with the microtubule for about 100 steps. In the cell, it is widely thought that several kinesin-1 motors cooperate to transport a cargo. Through a mechanistic model, we have extended the theoretical analysis of kinesin to describing transient and steady state behavior. A transient description is essential when studying collective behavior, as interaction between motors introduces time-varying loads. Herein, we interpret the kinesin motors as nonlinear, non-smooth oscillators and we employ metrics to characterize their cooperativity and to quantify their synchronization. These metrics are used to investigate the effect of the cargo linker stiffness, the load, and the difference in intrinsic velocity on the synchronization of two mechanically coupled motors.

1990 ◽  
Vol 22 (3) ◽  
pp. 564-586 ◽  
Author(s):  
Roland Malhamé

Electric water heating loads, in power systems, can be adequately modeled by Markov processes comprising a mix of continuous and discrete states. A physically-based characterization of the dynamic behavior of large aggregates of electric water heating loads is obtained by deriving the forward Kolmogorov equations associated with the individual hybrid-state processes. In addition, by focusing on the discrete part of the state, a Markov renewal viewpoint of the processes is developed. Both viewpoints are used to analyze and predict the transient and steady-state behavior of these loads, of great importance in load management applications.


1990 ◽  
Vol 22 (03) ◽  
pp. 564-586 ◽  
Author(s):  
Roland Malhamé

Electric water heating loads, in power systems, can be adequately modeled by Markov processes comprising a mix of continuous and discrete states. A physically-based characterization of the dynamic behavior of large aggregates of electric water heating loads is obtained by deriving the forward Kolmogorov equations associated with the individual hybrid-state processes. In addition, by focusing on the discrete part of the state, a Markov renewal viewpoint of the processes is developed. Both viewpoints are used to analyze and predict the transient and steady-state behavior of these loads, of great importance in load management applications.


Author(s):  
Saumya Yadav ◽  
Ambarish Kunwar

Molecular motor proteins are an extremely important component of the cellular transport system that harness chemical energy derived from ATP hydrolysis to carry out directed mechanical motion inside the cells. Transport properties of these motors such as processivity, velocity, and their load dependence have been well established through single-molecule experiments. Temperature dependent biophysical properties of molecular motors are now being probed using single-molecule experiments. Additionally, the temperature dependent biochemical properties of motors (ATPase activity) are probed to understand the underlying mechanisms and their possible implications on the enzymatic activity of motor proteins. These experiments in turn have revealed their activation energies and how they compare with the thermal energy available from the surrounding medium. In this review, we summarize such temperature dependent biophysical and biochemical properties of linear and rotary motor proteins and their implications for collective function during intracellular transport and cellular movement, respectively.


2013 ◽  
Vol 368 (1611) ◽  
pp. 20120023 ◽  
Author(s):  
Thomas Bilyard ◽  
Mayumi Nakanishi-Matsui ◽  
Bradley C. Steel ◽  
Teuta Pilizota ◽  
Ashley L. Nord ◽  
...  

The rotary motor F 1 -ATPase from the thermophilic Bacillus PS3 (TF 1 ) is one of the best-studied of all molecular machines. F 1 -ATPase is the part of the enzyme F 1 F O -ATP synthase that is responsible for generating most of the ATP in living cells. Single-molecule experiments have provided a detailed understanding of how ATP hydrolysis and synthesis are coupled to internal rotation within the motor. In this work, we present evidence that mesophilic F 1 -ATPase from Escherichia coli (EF 1 ) is governed by the same mechanism as TF 1 under laboratory conditions. Using optical microscopy to measure rotation of a variety of marker particles attached to the γ-subunit of single surface-bound EF 1 molecules, we characterized the ATP-binding, catalytic and inhibited states of EF 1 . We also show that the ATP-binding and catalytic states are separated by 35±3°. At room temperature, chemical processes occur faster in EF 1 than in TF 1 , and we present a methodology to compensate for artefacts that occur when the enzymatic rates are comparable to the experimental temporal resolution. Furthermore, we show that the molecule-to-molecule variation observed at high ATP concentration in our single-molecule assays can be accounted for by variation in the orientation of the rotating markers.


eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Ling Wang ◽  
Zachary Lee Johnson ◽  
Michael R Wasserman ◽  
Jesper Levring ◽  
Jue Chen ◽  
...  

ATP-binding cassette (ABC) transporters are molecular pumps ubiquitous across all kingdoms of life. While their structures have been widely reported, the kinetics governing their transport cycles remain largely unexplored. Multidrug resistance protein 1 (MRP1) is an ABC exporter that extrudes a variety of chemotherapeutic agents and native substrates. Previously, the structures of MRP1 were determined in an inward-facing (IF) or outward-facing (OF) conformation. Here, we used single-molecule fluorescence spectroscopy to track the conformational changes of bovine MRP1 (bMRP1) in real time. We also determined the structure of bMRP1 under active turnover conditions. Our results show that substrate stimulates ATP hydrolysis by accelerating the IF-to-OF transition. The rate-limiting step of the transport cycle is the dissociation of the nucleotide-binding-domain dimer, while ATP hydrolysis per se does not reset MRP1 to the resting state. The combination of structural and kinetic data illustrates how different conformations of MRP1 are temporally linked and how substrate and ATP alter protein dynamics to achieve active transport.


Author(s):  
Matthew J. Bovyn ◽  
Babu J.N. Reddy ◽  
Steven P. Gross ◽  
Jun F. Allard

AbstractMolecular motors like kinesin are critical for cellular organization and biological function including in neurons. There is detailed understanding of how they move and how factors such as applied force and the presence of microtubule-associated proteins can alter this single-motor travel. In order to walk, the cargo-motor complex must first attach to a microtubule. This attachment process is less studied. Here, we use a combination of single-molecule bead experiments, modeling, and simulation to examine how cargos with kinesin-1 bind to microtubules. In experiment, we find that increasing cargo size and environment viscosity both signficantly slow cargo binding time. We use modeling and simulation to examine how the single motor on rate translates to the on rate of the cargo. Combining experiment and modeling allows us to estimate the single motor on rate as 100 s−1. This is a much higher value than previous estimates. We attribute the difference between our measurements and previous estimates to two factors: first, we are directly measuring initial motor attachment (as opposed to re-binding of a second motor) and second, the theoretical framework allows us to account for missed events (i.e. binding events not detected by the experiments due to their short duration). This indicates that the mobility of the cargo itself, determined by its size and interaction with the cytoplasmic environment, play a previously underestimated role in determining intracellular transport kinetics.


2019 ◽  
Vol 47 (5) ◽  
pp. 1247-1257 ◽  
Author(s):  
Mateusz Dyla ◽  
Sara Basse Hansen ◽  
Poul Nissen ◽  
Magnus Kjaergaard

Abstract P-type ATPases transport ions across biological membranes against concentration gradients and are essential for all cells. They use the energy from ATP hydrolysis to propel large intramolecular movements, which drive vectorial transport of ions. Tight coordination of the motions of the pump is required to couple the two spatially distant processes of ion binding and ATP hydrolysis. Here, we review our current understanding of the structural dynamics of P-type ATPases, focusing primarily on Ca2+ pumps. We integrate different types of information that report on structural dynamics, primarily time-resolved fluorescence experiments including single-molecule Förster resonance energy transfer and molecular dynamics simulations, and interpret them in the framework provided by the numerous crystal structures of sarco/endoplasmic reticulum Ca2+-ATPase. We discuss the challenges in characterizing the dynamics of membrane pumps, and the likely impact of new technologies on the field.


Author(s):  
Fawzan Galib Abdul Karim Bawahab ◽  
Elvan Yuniarti ◽  
Edi Kurniawan

Abstrak. Pada penelitian ini, telah dilakukan analisa karakterisasi pada teknologi Direct Sequence Spread Spectrum dan Frequency Hopping Spread Spectrum, sebagai salah satu teknik multiple-access pada sistem komunikasi. Karakterisasi dilakukan untuk mencari bagaimana cara meningkatkan keoptimalan kedua sistem tersebut, dalam mengatasi masalah interferensi dengan sistem dan channel yang sama. Dan juga untuk menentukan veriabel apa yang mempengaruhi keoptimalan kedua sistem tersebut. Karakterisasi dilakukan dengan menentukan variabel-variabel yang mempengaruhi keoptimalan keduanya. Hasil dari karakterisasi, diketahui variabel-variabel yang mempengaruhi kemampuan sistem DSSS yaitu nilai frekuensi spreading (). Sedangkan untuk sistem FHSS yaitu nilai frekuensi spreading ( dan ) dan selisih antara frekuensi hopping data dengan frekuensi hopping interferensi . Kata Kunci: BER, DSSS, FHSS, Interference, Spread spectrum. Abstract. In this study, characterization of Direct Sequence Spread Spectrum and Frequency Hopping Spread Spectrum technologies have been done, as one of the multiple-access techniques in communication systems. Characterization is done to find out how to improve the ability of the two systems, in solving interference problems with the same system and channel. And also to determine what veriabel affects the ability of the two systems. Characterization is done by determining the variables that affect the ability of both. The results of the characterization, known variables that affect the ability of the DSSS system are the spreading frequency value (). As for the FHSS system, the spreading frequency value ( and ) and the difference between frequency hopping data with frequency hopping interference .


1984 ◽  
Vol 49 (2) ◽  
pp. 410-420
Author(s):  
Eva Hillerová ◽  
Miroslav Zdražil

Reversible adsorption of heptane and benzene on model and industrial hydrodesulphurization molybdena catalysts has been studied by elution chromatographic method at 150 °C. An increase in the adsorption of heptane on sulphidation of adsorbents was small for Al2O3 and great for MoO3. Supported catalysts behaved as mixture of Al2O3 and MoO3.The portion of surface which can be transformed by sulphidation into MoS2 ranged from 0 to 65% for individual commercial catalysts, as determined from the change in heptane adsorption after sulphidation of a given sample. The polarity of catalysts, including their acidity, was estimated from the difference between adsorption of benzene and heptane. The polarity of model and industrial catalysts in oxidic form was similar to that of alumina in most cases. The decrease in the polarity after sulphidation of the adsorbents was small for Al2O3 and great for MoO3. The decrease in polarity resulting from sulphidation of supported catalysts was relatively small, since the reaction of MoO3 monolayer with hydrogen sulphide leads to partial reformation of the alumina surface. The acidity of supported sulphided hydrodesulphurization catalysts has been shown by this method to be comparable with the acidity of the support itself.


2004 ◽  
Vol 82 (8) ◽  
pp. 1294-1303 ◽  
Author(s):  
Vanessa Renée Little ◽  
Keith Vaughan

1-Methylpiperazine was coupled with a series of diazonium salts to afford the 1-methyl-4-[2-aryl-1-diazenyl]piperazines (2), a new series of triazenes, which have been characterized by 1H and 13C NMR spectroscopy, IR spectroscopy, and elemental analysis. Assignment of the chemical shifts to specific protons and carbons in the piperazine ring was facilitated by comparison with the chemical shifts in the model compounds piperazine and 1-methylpiperazine and by a HETCOR experiment with the p-tolyl derivative (2i). A DEPT experiment with 1-methylpiperazine (6) was necessary to distinguish the methyl and methylene groups in 6, and a HETCOR spectrum of 6 enabled the correlation of proton and carbon chemical shifts. Line broadening of the signals from the ring methylene protons is attributed to restricted rotation around the N2-N3 bond of the triazene moiety in 2. The second series of triazenes, the ethyl 4-[2-phenyl-1-diazenyl]-1-piperazinecarboxylates (3), have been prepared by similar diazonium coupling to ethyl 1-piperazinecarboxylate and were similarly characterized. The chemical shifts of the piperazine ring protons are much closer together in series 3 than in series 2, resulting in distortion of the multiplets for these methylenes. It was noticed that the difference between these chemical shifts in 3 exhibited a linear free energy relationship with the Hammett substituent constants for the substituents in the aryl ring. Key words: triazene, piperazine, diazonium coupling, NMR, HETCOR, linear free energy relationship.


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