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Kinesin and ions

Junyu Xu, Na Wang, Jian-Hong Luo, Jun Xia
PICK1 (protein interacting with C-kinase 1) is a peripheral membrane protein that interacts with diverse membrane proteins. PICK1 has been shown to regulate the clustering and membrane localization of synaptic receptors such as AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptors, metabotropic glutamate receptor 7, and ASICs (acid-sensing ion channels). Moreover, recent evidence suggests that PICK1 can mediate the trafficking of various vesicles out from the Golgi complex in several cell systems, including neurons...
2016: Scientific Reports
Amber L Jolly, Chi-Hao Luan, Brendon E Dusel, Sara F Dunne, Michael Winding, Vishrut J Dixit, Chloe Robins, Jennifer L Saluk, David J Logan, Anne E Carpenter, Manu Sharma, Deborah Dean, Andrew R Cohen, Vladimir I Gelfand
Long-distance intracellular transport of organelles, mRNA, and proteins ("cargo") occurs along the microtubule cytoskeleton by the action of kinesin and dynein motor proteins, but the vast network of factors involved in regulating intracellular cargo transport are still unknown. We capitalize on the Drosophila melanogaster S2 model cell system to monitor lysosome transport along microtubule bundles, which require enzymatically active kinesin-1 motor protein for their formation. We use an automated tracking program and a naive Bayesian classifier for the multivariate motility data to analyze 15,683 gene phenotypes and find 98 proteins involved in regulating lysosome motility along microtubules and 48 involved in the formation of microtubule filled processes in S2 cells...
January 26, 2016: Cell Reports
Udo Rüb, Horst-Werner Korf
Neurons are extremely polarized cells with a complex and unique morphology, as well as long processes (axon and dendrites) that extend far away from the nerve cell body. This architectonic feature of polarization with afferent (i.e., dendrites) and efferent processes (i.e., axons) requires efficient communication between the body of nerve cells and their periphery and makes nerve cells particularly dependent on functionally intact, sufficient, and timely axonal and/or dendritic intracellular transport processes over long distances...
2015: Advances in Anatomy, Embryology, and Cell Biology
Hui-Ming Zhang, Simon Wheeler, Xue Xia, Ruslana Radchuk, Hans Weber, Christina E Offler, John W Patrick
BACKGROUND: Transfer cells are characterized by intricate ingrowth walls, comprising an uniform wall upon which wall ingrowths are deposited. The ingrowth wall forms a scaffold to support an amplified plasma membrane surface area enriched in membrane transporters that collectively confers transfer cells with an enhanced capacity for membrane transport at bottlenecks for apo-/symplasmic exchange of nutrients. However, the underlying molecular mechanisms regulating polarized construction of the ingrowth wall and membrane transporter profile are poorly understood...
2015: BMC Plant Biology
Lian Duan, Tong-Qing Wang, Wei Bian, Wen Liu, Yue Sun, Bin-Sheng Yang
Monastrol, a cell-permeable inhibitor, considered to specifically inhibit kinesin Eg5, can cause mitotic arrest and monopolar spindle formation, thus exhibiting antitumor properties. Centrin, a ubiquitous protein associated with centrosome, plays a critical role in centrosome duplication. Moreover, a correlation between centrosome amplification and cancer has been reported. In this study, it is proposed for the first time that centrin may be another target of the anticancer drug monastrol since monastrol can effectively inhibit not only the growth of the transformed Escherichia coli cells in vivo, but also the Lu(3+)-dependent self-assembly of EoCen in vitro...
February 25, 2015: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
Stefan Dhein, Anna Schreiber, Sabine Steinbach, Daniel Apel, Aida Salameh, Franziska Schlegel, Martin Kostelka, Pascal M Dohmen, Friedrich Wilhelm Mohr
OBJECTIVES: The aim of our study was to elucidate how cyclic mechanical stretch is sensed by cardiomyocytes and in which way it affects cytoskeletal organization. METHODS: Neonatal rat cardiomyocytes, cultured on flexible membranes, were subjected to cyclic mechanical stretch (1 Hz, 10% elongation) for 24 h using either round or rectangular loading posts for equibi-axial or uni-axial stretch, respectively, using the FlexCell stretch system. Cells were treated either with vehicle, the focal adhesion kinase (FAK) inhibitor PF-573,228 (200 nM), or the stretch-activated ion channel blocker gadolinium (Gd(3+); 100 μM)...
August 2014: Progress in Biophysics and Molecular Biology
Konrad J Böhm
Copper is a trace element required to maintain essential life processes. In healthy organisms, copper metabolism is well balanced. If this balance is destroyed, the cellular level of free copper might increase and cause toxic effects. So far, the molecular mechanisms of copper intoxication are understood only partly. The present study revealed that the kinesin-dependent transport system is strongly affected by copper(II) ions. Both the microtubules, along which kinesin moves, and the kinesin itself were found to be the target structures of copper ions: Microtubule formation was suppressed by copper ions (IC50 26-70 µM) apparently chiefly by inhibition of binding of microtubule-associated proteins to tubulin...
April 2015: Archives of Toxicology
Christopher I Ugbode, Warren D Hirst, Marcus Rattray
Recent evidence suggests that the predominant astrocyte glutamate transporter, GLT-1/ Excitatory Amino Acid Transporter 2 (EAAT2) is associated with mitochondria. We used primary cultures of mouse astrocytes to assess co-localization of GLT-1 with mitochondria, and tested whether the interaction was dependent on neurons, actin polymerization or the kinesin adaptor, TRAK2. Mouse primary astrocytes were transfected with constructs expressing V5-tagged GLT-1, pDsRed1-Mito with and without dominant negative TRAK2...
September 2014: Journal of Neurochemistry
Konrad J Böhm
The anterograde vesicle transport within neurons critically depends on microtubules and on the activity of kinesin. The present study demonstrates that cadmium ions inhibit the in vitro assembly of microtubules from tubulin, whereby at high cadmium levels (∼500 μM) unstructured protein aggregates were formed. Cadmium ions also significantly lower both the ATPase and motility activity of neuron-specific kinesin KIF5A in concentration-dependent manner. For the inhibition of KIF5A ATPase activity, an IC50 value of 10...
January 30, 2014: Toxicology Letters
Philip Klepeisz, Sandra Sagmeister, Verena Haudek-Prinz, Melanie Pichlbauer, Bettina Grasl-Kraupp, Christopher Gerner
Preceding studies on the mode of action of non-genotoxic hepatocarcinogens (NGCs) have concentrated on alterations induced in hepatocytes (HCs). A potential role of non-parenchymal liver cells (NPCs) in NGC-driven hepatocarcinogenesis has been largely neglected so far. The aim of this study is to characterize NGC-induced alterations in the proteome profiles of HCs as well as NPCs. We chose the prototypic NGC phenobarbital (PB) which was applied to male rats for a period of 14 days. The livers of PB-treated rats were perfused by collagenase and the cell suspensions obtained were subjected to density gradient centrifugation to separate HCs from NPCs...
2013: PloS One
Helmut Plattner
The contractile vacuole complex (CVC) of freshwater protists sequesters the excess of water and ions (Ca(2+)) for exocytosis cycles at the pore. Sequestration is based on a chemiosmotic proton gradient produced by a V-type H(+)-ATPase. So far, many pieces of information available have not been combined to a comprehensive view on CVC biogenesis and function. One main function now appears as follows. Ca(2+)-release channels, type inositol 1,4,5-trisphosphate receptors (InsP3R), may serve for fine-tuning of local cytosolic Ca(2+) concentration and mediate numerous membrane-to-membrane interactions within the tubular spongiome meshwork...
June 2015: Critical Reviews in Microbiology
Hitomi Mitsugi, Takeshi Niki, Kazuko Takahashi-Niki, Kyoko Tanimura, Kumiko Yoshizawa-Kumagaye, Masahiko Tsunemi, Sanae M M Iguchi-Ariga, Hiroyoshi Ariga
DJ-1, the product of familial Parkinson's disease gene and an oncogene, is a cysteine protease which plays a role in anti-oxidative stress reaction. In this study, we identified the recognition sequence for DJ-1 protease by using recombinant DJ-1 and a peptide library. Protease activity of DJ-1 lacking C-terminal α-helix (DJ-1ΔH9) was stronger than that of full-sized DJ-1, and the most susceptible sequence digested by DJ-1ΔH9 was valine-lysine-valine-alanine (VKVA) under the optimal conditions of pH 5.5 and 0 mM NaCl...
August 19, 2013: FEBS Letters
Noriyuki Uchida, Kou Okuro, Yamato Niitani, Xiao Ling, Takayuki Ariga, Michio Tomishige, Takuzo Aida
A water-soluble dendron with a fluorescein isothiocyanate (FITC) fluorescent label and bearing nine pendant guanidinium ion (Gu(+))/benzophenone (BP) pairs at its periphery (Glue(BP)-FITC) serves as a "photoclickable molecular glue". By multivalent salt-bridge formation between Gu(+) ions and oxyanions, Glue(BP)-FITC temporarily adheres to a kinesin/microtubule hybrid. Upon subsequent exposure to UV light, this noncovalent binding is made permanent via a cross-linking reaction mediated by carbon radicals derived from the photoexcited BP units...
March 27, 2013: Journal of the American Chemical Society
Christian J Koehler, Magnus Ø Arntzen, Gustavo Antonio de Souza, Bernd Thiede
Isobaric peptide termini labeling (IPTL) is based on labeling of both peptide termini with complementary isotopic labels resulting in isobaric peptides. MS/MS analysis after IPTL derivatization produces peptide-specific fragment ions which are distributed throughout the MS/MS spectrum. Thus, several quantification points can be obtained per peptide. In this report, we present triplex-IPTL, a chemical labeling strategy for IPTL allowing the simultaneous quantification of three states within one MS run. For this purpose, dimethylation of the N-terminal amino group followed by dimethylation of lysines was used with different stable isotopes of formaldehyde and cyanoborohydride...
February 19, 2013: Analytical Chemistry
Slobodanka Korten, Nuria Albet-Torres, Francesca Paderi, Lasse ten Siethoff, Stefan Diez, Till Korten, Geertruy te Kronnie, Alf Månsson
The last decade has seen appreciable advancements in efforts towards increased portability of lab-on-a-chip devices by substituting microfluidics with molecular motor-based transportation. As of now, first proof-of-principle devices have analyzed protein mixtures of low complexity, such as target protein molecules in buffer solutions optimized for molecular motor performance. However, in a diagnostic work-up, lab-on-a-chip devices need to be compatible with complex biological samples. While it has been shown that such samples do not interfere with crucial steps in molecular diagnostics (for example antibody-antigen recognition), their effect on molecular motors is unknown...
March 7, 2013: Lab on a Chip
Maike H Hinrichs, Avesta Jalal, Bernhard Brenner, Eckhard Mandelkow, Satish Kumar, Tim Scholz
Current models for the intracellular transport of Tau protein suggest motor protein-dependent co-transport with microtubule fragments and diffusion of Tau in the cytoplasm, whereas Tau is believed to be stationary while bound to microtubules and in equilibrium with free diffusion in the cytosol. Observations that members of the microtubule-dependent kinesin family show Brownian motion along microtubules led us to hypothesize that diffusion along microtubules could also be relevant in the case of Tau. We used single-molecule total internal reflection fluorescence microscopy to probe for diffusion of individual fluorescently labeled Tau molecules along microtubules...
November 9, 2012: Journal of Biological Chemistry
Joshua Barry, Chen Gu
Proper localization of various ion channels is fundamental to neuronal functions, including postsynaptic potential plasticity, dendritic integration, action potential initiation and propagation, and neurotransmitter release. Microtubule-based forward transport mediated by kinesin motors plays a key role in placing ion channel proteins to correct subcellular compartments. PDZ- and coiled-coil-domain proteins function as adaptor proteins linking ionotropic glutamate and GABA receptors to various kinesin motors, respectively...
April 2013: Neuroscientist: a Review Journal Bringing Neurobiology, Neurology and Psychiatry
Elena V Tsimakouridze, Marty Straume, Peter S Podobed, Heather Chin, Jonathan LaMarre, Ron Johnson, Monica Antenos, Gordon M Kirby, Allison Mackay, Patsy Huether, Jeremy A Simpson, Michael Sole, Gerard Gadal, Tami A Martino
There is critical demand in contemporary medicine for gene expression markers in all areas of human disease, for early detection of disease, classification, prognosis, and response to therapy. The integrity of circadian gene expression underlies cardiovascular health and disease; however time-of-day profiling in heart disease has never been examined. We hypothesized that a time-of-day chronomic approach using samples collected across 24-h cycles and analyzed by microarrays and bioinformatics advances contemporary approaches, because it includes sleep-time and/or wake-time molecular responses...
August 2012: Chronobiology International
Xinghua Qin, Ziwei Chen, Tao Xu, Ping Li, Guoqin Liu
GhKCH2, a member of the kinesin superfamily, is a plant-specific microtubule-dependent motor protein from cotton with the ability to bind to both microtubules and microfilaments. Here, the motor domain of GhKCH2 (GhKCH2MD; amino acids 371-748) was overexpressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method. The pH of the crystallization buffer was shown to have a significant effect on the crystal morphology and diffraction quality. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 60...
July 1, 2012: Acta Crystallographica. Section F, Structural Biology and Crystallization Communications
Jared C Cochran, Yu Cheng Zhao, Dean E Wilcox, F Jon Kull
Kinesins are molecular motors that require a divalent metal ion (for example, Mg(2+)) to convert the energy of ATP hydrolysis into directed force production along microtubules. Here we present the crystal structure of a recombinant kinesin motor domain bound to Mn(2+) and ADP and report on a serine-to-cysteine substitution in the switch 1 motif of kinesin that allows its ATP hydrolysis activity to be controlled by adjusting the ratio of Mn(2+) to Mg(2+). This mutant kinesin binds ATP similarly in the presence of either metal ion, but its ATP hydrolysis activity is greatly diminished in the presence of Mg(2+)...
January 2012: Nature Structural & Molecular Biology
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