Localization of neuronal growth-associated, microtubule-destabilizing factor SCG10 in brain-derived raft membrane microdomains.
Raft is a mobile membrane subdomain enriched in sphingolipid and cholesterol and also various signaling molecules. Previous observation suggested that brain-derived rafts contain tubulin but that rafts of non-neural origin do not. We hypothesized that SCG10, one of the neuronal growth-associated proteins (nGAPs), might be a neuron-specific molecule that anchors tubulin to neuronal rafts, and we explored biochemically its subcellular localization, interaction with tubulin, and effects on microtubule dynamics. In postnatal rat brain extracts, SCG10 was recovered mostly in membrane-associated fractions, and at least half was included in the raft fraction that was also enriched in GAP-43 and NAP-22. SCG10-enriched brain rafts also contained tubulin, and chemical cross-linking experiments revealed that SCG10 was closely associated with tubulin. In addition, SCG10 was able to inhibit polymerization of tubulin. These results indicate that SCG10 is a component of neuronal rafts as are other nGAPs, and suggest that SCG10 may be involved in signaling events in membranes for cytoskeletal reorganization around neuronal rafts.
Maekawa
S
S
Department of Applied Biology, Kyoto Institute of Technology, Kyoto 606-8585, Japan. morinosm@nils.go.jp
Morii
H
H
Kumanogoh
H
H
Sano
M
M
Naruse
Y
Y
Sokawa
Y
Y
Mori
N
N
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002352:Carrier Proteins; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D021962:Membrane Microdomains / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins; D008868:Microtubule Proteins; D008870:Microtubules / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D009414:Nerve Growth Factors / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D009474:Neurons / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000502:physiology; D013347:Subcellular Fractions / Q000737:chemistry; D014404:Tubulin / Q000737:chemistry*
D002352:Carrier Proteins; D008565:Membrane Proteins; D008868:Microtubule Proteins; D009414:Nerve Growth Factors; C512848:Stmn2 protein, rat; D014404:Tubulin