Linked Life Data

11012681

Source:http://linkedlifedata.com/resource/pubmed/id/11012681

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
20
pubmed:dateCreated
2000-11-28
pubmed:abstractText
The association of membrane-bounded cell organelles to microtubules is crucial for determination of their shape, intracellular localization and translocation. We have shown previously the high affinity binding of peroxisomes to microtubules which appears to be of static nature as in vivo studies indicate that only a few peroxisomes move along the microtubular tracks. In order to characterize the interactions of peroxisomes with microtubules, we have developed a semiquantitative in vitro binding assay, which is based on the association of highly purified rat liver peroxisomes to microtubules coated onto microtiterplates. The binding was visualized by differential interference contrast and immunofluorescence using a confocal laser scanning microscope. The binding was concentration dependent and saturable, being affected by time, temperature, and pH. Addition of ATP or the motor proteins kinesin and dynein increased the binding capacity, while ATP-depletion or microtubule associated proteins (MAPs) decreased it. KCl treatment of peroxisomes reduced the binding, which was restored by dialyzed KCl-stripping eluate as well as by rat liver cytosol. The reconstituting effect of cytosol was abolished by its pretreatment with proteases or N-ethylmaleimide. Moreover, the treatment of peroxisomes with proteases or N-ethylmaleimide reduced their binding, which was not reversed by cytosol. These results suggest the involvement of a peroxisomal membrane protein and cytosolic factor(s) in the binding of peroxisomes to microtubules. This notion is supported by the observation that distinct subfractions of dialyzed KCl-stripping eluate obtained by gel chromatography augmented the binding. Those subfractions, as well as purified peroxisome fractions, exhibited strong immunoreactivity with an antibody to cytoplasmic linker protein (CLIP)-115, revealing a 70-kDa polypeptide. Moreover, immunodepletion of KCl-stripping eluate and its subfractions with an antibody to the conserved microtubule binding domain of CLIPs, abolished their promoting effect on the binding, thus suggesting the involvement of a CLIP-related protein in the binding of peroxisomes to microtubules.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0014-2956
pubmed:author
pubmed:issnType
Print
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
6264-75
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:11012681-Adenosine Triphosphate, pubmed-meshheading:11012681-Animals, pubmed-meshheading:11012681-Cell Fractionation, pubmed-meshheading:11012681-Cell Membrane, pubmed-meshheading:11012681-Cytosol, pubmed-meshheading:11012681-Dyneins, pubmed-meshheading:11012681-Ethylmaleimide, pubmed-meshheading:11012681-Female, pubmed-meshheading:11012681-Hydrogen-Ion Concentration, pubmed-meshheading:11012681-Kinesin, pubmed-meshheading:11012681-Kinetics, pubmed-meshheading:11012681-Liver, pubmed-meshheading:11012681-Microscopy, Confocal, pubmed-meshheading:11012681-Microtubule-Associated Proteins, pubmed-meshheading:11012681-Microtubules, pubmed-meshheading:11012681-Models, Biological, pubmed-meshheading:11012681-Peroxisomes, pubmed-meshheading:11012681-Potassium Chloride, pubmed-meshheading:11012681-Rats, pubmed-meshheading:11012681-Rats, Sprague-Dawley, pubmed-meshheading:11012681-Thermodynamics
pubmed:year
2000
pubmed:articleTitle
Interaction of peroxisomes with microtubules. In vitro studies using a novel peroxisome-microtubule binding assay.
pubmed:affiliation
Department of Anatomy and Cell Biology, Division of Medical Cell Biology, University of Heidelberg, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't