Fast axonal transport of kinesin in the rat visual system: functionality of kinesin heavy chain isoforms.
The mechanochemical ATPase kinesin is thought to move membrane-bounded organelles along microtubules in fast axonal transport. However, fast transport includes several classes of organelles moving at rates that differ by an order of magnitude. Further, the fact that cytoplasmic forms of kinesin exist suggests that kinesins might move cytoplasmic structures such as the cytoskeleton. To define cellular roles for kinesin, the axonal transport of kinesin was characterized. Retinal proteins were pulse-labeled, and movement of radiolabeled kinesin through optic nerve and tract into the terminals was monitored by immunoprecipitation. Heavy and light chains of kinesin appeared in nerve and tract at times consistent with fast transport. Little or no kinesin moved with slow axonal transport indicating that effectively all axonal kinesin is associated with membranous organelles. Both kinesin heavy chain molecular weight variants of 130,000 and 124,000 M(r) (KHC-A and KHC-B) moved in fast anterograde transport, but KHC-A moved at 5-6 times the rate of KHC-B. KHC-A cotransported with the synaptic vesicle marker synaptophysin, while a portion of KHC-B cotransported with the mitochondrial marker hexokinase. These results suggest that KHC-A is enriched on small tubulovesicular structures like synaptic vesicles and that at least one form of KHC-B is predominantly on mitochondria. Biochemical specialization may target kinesins to appropriate organelles and facilitate differential regulation of transport.
Elluru
R G
RG
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235-9111, USA.
Bloom
G S
GS
Brady
S T
ST
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001370:Axonal Transport; D002451:Cell Compartmentation; D005136:Eye Proteins / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D016547:Kinesins / Q000737:chemistry / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D008970:Molecular Weight; D009897:Optic Chiasm / Q000378:metabolism; D009900:Optic Nerve / Q000378:metabolism*; D015388:Organelles / Q000378:metabolism; D011233:Precipitin Tests; D011487:Protein Conformation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism; D016708:Synaptophysin / Q000378:metabolism; D013997:Time Factors; D014795:Visual Pathways / Q000378:metabolism
D000911:Antibodies, Monoclonal; D005136:Eye Proteins; D007527:Isoenzymes; D016708:Synaptophysin; D006593:Hexokinase; D016547:Kinesins