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Palmitoylcarnitine Affects Localization of Growth Associated Protein GAP-43 in Plasma Membrane Subdomains and its Interaction with Gα$_{o}$ in Neuroblastoma NB-2a Cells

Content Provider	Springer Nature Link
Author	Tułodziecka, Karolina Czeredys, Magdalena Nałęcz, Katarzyna A.
Copyright Year	2012
Abstract	Palmitoylcarnitine was observed previously to promote differentiation of neuroblastoma NB-2a cells, and to affect protein kinase C (PKC). Palmitoylcarnitine was also observed to increase palmitoylation of several proteins, including a PKC substrate, whose expression augments during differentiation of neural cells—a growth associated protein GAP-43, known to bind phosphatidylinositol 4,5-bisphosphate [PI(4,5)P$_{2}$]. Since palmitoylated proteins are preferentially localized in sphingolipid- and cholesterol-rich microdomains of plasma membrane, the present study has been focused on a possible effect of palmitoylcarnitine on GAP-43 localization in these microdomains. Palmitoylcarnitine treatment resulted in GAP-43 appearance in floating fractions (rafts) in sucrose gradient and increased co-localization with cholesterol and with PI(4,5)P$_{2}$, although co-localization of both lipids decreased. GAP-43 disappeared from raft fraction upon treatment with 2-bromopalmitate (an inhibitor of palmitoylating enzymes) and after treatment with etomoxir (carnitine palmitoyltransferase I inhibitor). Raft localization of GAP-43 was completely abolished by treatment with methyl-β-cyclodextrin, a cholesterol binding agent, while there was no change upon sequestration of PI(4,5)P$_{2}$ with neomycin. GAP-43 co-precipitated with a monomeric form of Gα$_{o}$, a phenomenon diminished after palmitoylcarnitine treatment and paralleled by a decrease of Gα$_{o}$ in the raft fraction. These observations point to palmitoylation of GAP-43 as a mechanism leading to an increased localization of this protein in microdomains of plasma membrane rich in cholesterol, in majority different, however, from microdomains in which PI(4,5)P$_{2}$ is present. This localization correlates with decreased interaction with Gα$_{o}$ and suppression of its activity—an important step regulating neural cell differentiation.
Starting Page	519
Ending Page	529
Page Count	11
File Format	PDF
ISSN	03643190
Journal	Neurochemical Research
Volume Number	38
Issue Number	3
e-ISSN	15736903
Language	English
Publisher	Springer US
Publisher Date	2012-12-09
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Palmitoylcarnitine GAP-43 Cholesterol Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P$_{2}$] Gα$_{o}$ Neurosciences Neurochemistry Biochemistry Cell Biology Neurology
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Cellular and Molecular Neuroscience

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Palmitoylcarnitine Affects Localization of Growth Associated Protein GAP-43 in Plasma Membrane Subdomains and its Interaction with Gα$_{o}$ in Neuroblastoma NB-2a Cells