Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0012739 (disseminated intravascular coagulation)
 8,673 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To facilitate functional studies of novel myosins, we have developed a strategy for characterizing the mechanochemical properties of motors isolated by immunoadsorption directly from small amounts of crude tissue extracts. In this initial study, silica beads coated with an antibody that specifically recognizes the tail of myosin-V were used to immunoadsorb this motor protein from brain extracts. The myosin-containing beads were then positioned with optical tweezers onto actin filaments nucleated from Limulus sperm acrosomal processes and observed for motility using high resolution video DIC microscopy. The addition of brush border spectrin to the motility chamber enabled the growth of stable actin filament tracks that were approximately 4-fold longer than filaments grown in the absence of this actin crosslinking protein. The velocity of myosin-V immunoadsorbed from brain extracts was similar to that observed for purified myosin-V that was antibody-linked to beads or assessed using the sliding actin filament assay. Motile beads containing myosin-V immunoadsorbed from brain extracts bound poorly to nucleated actin filaments and were incapable of linear migrations following the addition of a different antibody that specifically recognizes the motor-containing head domain of myosin-V. Myosin-V motility was most robust in the absence of Ca2+. Interestingly, skeletal muscle tropomyosin and brush border spectrin had no detectable effect on myosin-V mechanochemistry. Myosin-V containing beads were also occasionally observed migrating directly on acrosomal processes in the absence of exogenously added actin.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:In vitro motility of immunoadsorbed brain myosin-V using a Limulus acrosomal process and optical tweezer-based assay. 761 69

Active neuronal transport along microtubules participates in the targeting of mRNAs, proteins and organelles to their sites of action. Cytoplasmic dynein represents a minus-end-directed microtubule-dependent motor protein. Due to the polarity of microtubules in axonal and distal dendritic compartments, with microtubule minus-ends pointing toward the inside of the cell, dyneins mainly mediate retrograde transport pathways in neurons. Since dyneins transport synaptic proteins, we asked whether changes in neuronal activity would in general influence dynein transport. KCl-induced depolarization, a condition that mimics the effects of neuronal activity, or pharmacological blockade of neuronal action potentials, respectively, was combined with neuronal live cell imaging, using an autofluorescent dynein intermediate chain fusion (monomeric red fluorescent protein [mRFP]-dynein intermediate chain [DIC]) as a model protein. Notably, we found that induced activity significantly reduced dynein particle mobility, as well as both the total distance and velocity of movements in mouse cultured hippocampal neurons. In contrast, blockade of neuronal action potentials through TTX did not alter any of the parameters analyzed. Neuronal depolarization processes therefore represent candidate mechanisms to regulate intracellular transport of neuronal cargoes.
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PMID:Neuronal depolarization modifies motor protein mobility. 1925 Sep 60