TY - JOUR
T1 - A novel split kinesin assay identifies motor proteins that interact with distinct vesicle populations
AU - Jenkins, Brian
AU - Decker, Helena
AU - Bentley, Marvin
AU - Luisi, Julie
AU - Banker, Gary
PY - 2012/8
Y1 - 2012/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84865978266&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84865978266&partnerID=8YFLogxK
U2 - 10.1083/jcb.201205070
DO - 10.1083/jcb.201205070
M3 - Article
C2 - 22908316
AN - SCOPUS:84865978266
SN - 0021-9525
VL - 198
SP - 749
EP - 761
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 4
ER -