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Compartmentalization of membrane proteins by the actin cytoskeleton

dc.contributor.authorHiggins, Jenny, author
dc.contributor.authorKrapf, Diego, advisor
dc.contributor.authorTamkun, Michael, committee member
dc.contributor.authorBamburg, James, committee member
dc.contributor.authorAzimi-Sadjadi, Mahmood, committee member
dc.date.accessioned2007-01-03T06:11:26Z
dc.date.available2015-01-31T05:57:00Z
dc.date.issued2013
dc.description.abstractActing as the point of contact for the outside world, the plasma membrane is crucial for cellular signaling events. Proper organization of membrane components is necessary to accomplish this task. Although a number of experiments have demonstrated the compartmentalization of lipids and proteins on the plasma membrane, direct observation of the mechanisms by which the organization occurs has been challenging, in part due to the imaging restrictions of a diffraction-limited system and the dynamic nature of the membrane compartmentalization. Using photoactivated localization microscopy (PALM), a superresolution technique, we have captured the dynamics of compartments formed by the cortical actin cytoskeleton. Live human embryonic kidney (HEK293) cells were imaged with a temporal resolution of 2 s and a spatial resolution of 40 nm. The actin cytoskeleton forms compartments with a mean area of 2.3±0.3 μm2 that are partially outlined by actin bundles. When the PALM images of actin were combined with single particle tracking of membrane proteins, we directly observed the cytoskeleton acting as a barrier to the diffusion of Kv2.1 and Kv1.4, two voltage-gated potassium channels. In addition, we used a novel compartment detection and tracking algorithm to show that Kv2.1 and Kv1.4 channels avoid actin when changing compartments. This work represents the first direct observations of individual membrane protein interactions with barriers formed by the actin cytoskeleton.
dc.format.mediumborn digital
dc.format.mediummasters theses
dc.identifierHiggins_colostate_0053N_12048.pdf
dc.identifierETDF2013500381BIOM
dc.identifier.urihttp://hdl.handle.net/10217/81011
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2000-2019
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.subjectmembrane protein
dc.subjectsuperresolution
dc.subjectplasma membrane compartmentalization
dc.subjectactin
dc.subjectcytoskeleton
dc.subjectPALM
dc.titleCompartmentalization of membrane proteins by the actin cytoskeleton
dc.typeText
dcterms.embargo.expires2015-01-31
dcterms.embargo.terms2015-01-31
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineBiomedical Engineering
thesis.degree.grantorColorado State University
thesis.degree.levelMasters
thesis.degree.nameMaster of Science (M.S.)

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