A novel split kinesin assay identifies motor proteins that interact with distinct vesicle populations

Brian Jenkins, Helena Decker, Marvin Bentley, Julie Luisi, Gary Banker

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

Identifying the kinesin motors that interact with different vesicle populations is a longstanding and challenging problem with implications for many aspects of cell biology. Here we introduce a new live-cell assay to assess kinesin-vesicle interactions and use it to identify kinesins that bind to vesicles undergoing dendrite-selective transport in cultured hippocampal neurons. We prepared a library of "split kinesins," comprising an axon-selective kinesin motor domain and a series of kinesin tail domains that can attach to their native vesicles; when the split kine-sins were assembled by chemical dimerization, bound vesicles were misdirected into the axon. This method provided highly specific results, showing that three Kinesin-3 family members-KIF1A, KIF13A, and KIF13B-interacted with dendritic vesicle populations. This experimental paradigm allows a systematic approach to evaluate motor-vesicle interactions in living cells.

Original languageEnglish (US)
Pages (from-to)749-761
Number of pages13
JournalJournal of Cell Biology
Volume198
Issue number4
DOIs
StatePublished - Aug 1 2012

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ASJC Scopus subject areas

  • Cell Biology

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