Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells

Kinesin motor proteins drive the transport of cellular cargoes along microtubule tracks. How motor protein activity is controlled in cells is unresolved, but it is likely coupled to changes in protein conformation and cargo association. By applying the quantitative method fluorescence resonance ener...

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Main Authors:	Cai, Dawen, Hoppe, Adam D., Swanson, Joel A., Verhey, Kristen J.
Format:	Online
Language:	English
Published:	The Rockefeller University Press 2007
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2063625/

id	pubmed-2063625
recordtype	oai_dc
spelling	pubmed-20636252007-11-29 Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells Cai, Dawen Hoppe, Adam D. Swanson, Joel A. Verhey, Kristen J. Research Articles Kinesin motor proteins drive the transport of cellular cargoes along microtubule tracks. How motor protein activity is controlled in cells is unresolved, but it is likely coupled to changes in protein conformation and cargo association. By applying the quantitative method fluorescence resonance energy transfer (FRET) stoichiometry to fluorescent protein (FP)–labeled kinesin heavy chain (KHC) and kinesin light chain (KLC) subunits in live cells, we studied the overall structural organization and conformation of Kinesin-1 in the active and inactive states. Inactive Kinesin-1 molecules are folded and autoinhibited such that the KHC tail blocks the initial interaction of the KHC motor with the microtubule. In addition, in the inactive state, the KHC motor domains are pushed apart by the KLC subunit. Thus, FRET stoichiometry reveals conformational changes of a protein complex in live cells. For Kinesin-1, activation requires a global conformational change that separates the KHC motor and tail domains and a local conformational change that moves the KHC motor domains closer together. The Rockefeller University Press 2007-01-01 /pmc/articles/PMC2063625/ /pubmed/17200416 http://dx.doi.org/10.1083/jcb.200605097 Text en Copyright © 2007, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Cai, Dawen Hoppe, Adam D. Swanson, Joel A. Verhey, Kristen J.
spellingShingle	Cai, Dawen Hoppe, Adam D. Swanson, Joel A. Verhey, Kristen J. Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
author_facet	Cai, Dawen Hoppe, Adam D. Swanson, Joel A. Verhey, Kristen J.
author_sort	Cai, Dawen
title	Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
title_short	Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
title_full	Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
title_fullStr	Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
title_full_unstemmed	Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells
title_sort	kinesin-1 structural organization and conformational changes revealed by fret stoichiometry in live cells
description	Kinesin motor proteins drive the transport of cellular cargoes along microtubule tracks. How motor protein activity is controlled in cells is unresolved, but it is likely coupled to changes in protein conformation and cargo association. By applying the quantitative method fluorescence resonance energy transfer (FRET) stoichiometry to fluorescent protein (FP)–labeled kinesin heavy chain (KHC) and kinesin light chain (KLC) subunits in live cells, we studied the overall structural organization and conformation of Kinesin-1 in the active and inactive states. Inactive Kinesin-1 molecules are folded and autoinhibited such that the KHC tail blocks the initial interaction of the KHC motor with the microtubule. In addition, in the inactive state, the KHC motor domains are pushed apart by the KLC subunit. Thus, FRET stoichiometry reveals conformational changes of a protein complex in live cells. For Kinesin-1, activation requires a global conformational change that separates the KHC motor and tail domains and a local conformational change that moves the KHC motor domains closer together.
publisher	The Rockefeller University Press
publishDate	2007
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2063625/
_version_	1611406173144088576

Kinesin-1 structural organization and conformational changes revealed by FRET stoichiometry in live cells

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