title: Neuroglobin promotes neurite outgrowth via differential binding to PTEN and Akt.
abstract: Neuroglobin, the third mammalian globin with a hexa-coordinated heme, exists predominantly in neurons of the brain. Neuroglobin plays an important role in neuronal death upon ischemia and oxidative stress. The physiological function of neuroglobin remains unclear. Here, we report a novel function of neuroglobin in neurite development. Knocking-down neuroglobin exhibited a prominent neurite-deficient phenotype in mouse neuroblastoma N2a cells. Silencing neuroglobin prevented neurite outgrowth, while ectopic expression of neuroglobin but not homologous cytoglobin promoted neurite outgrowth of N2a cells upon serum withdrawal. In primary cultured rat cerebral cortical neurons, neuroglobin was upregulated and preferentially distributed in neurites during neuronal development. Overexpression of neuroglobin but not cytoglobin in cultured cortical neurons promoted axonal outgrowth, while knocking-down of neuroglobin retarded axonal outgrowth. Neuroglobin overexpression suppressed phosphatase and tensin homolog (PTEN) but increased Akt phosphorylation during neurite induction. Bimolecular fluorescence complementation and glutathione S-transferase pull-down assays revealed that neuroglobin and various mutants (E53Q, E118Q, K119N, H64A, H64L, and Y44D) bound with Akt and PTEN differentially. Neuroglobin E53Q showed a prominent reduced PTEN binding but increased Akt binding, resulting in decreased p-PTEN, increased p-Akt, and increased neurite length. Taken together, we demonstrate a critical role of neuroglobin in neuritogenesis or development via interacting with PTEN and Akt differentially to activate phosphatidylinositol 3-kinase/Akt pathway, providing potential therapeutic applications of neuroglobin for axonopathy in neurological diseases.
authors.affiliation: School of Basic Medicine, Tongji Medical College, Wuhan, China.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Qian Rong
authors.initials: QR
authors.lastname: Liu
authors.forename: Xin Xin
authors.initials: XX
authors.lastname: Xiong
authors.forename: Ju Mei
authors.initials: JM
authors.lastname: Liu
authors.forename: Xiao Jing
authors.initials: XJ
authors.lastname: Lai
authors.forename: Chun
authors.initials: C
authors.lastname: Cheng
authors.forename: Feng
authors.initials: F
authors.lastname: Pan
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Shang Bin
authors.initials: SB
authors.lastname: Yu
authors.forename: Albert Cheung Hoi
authors.initials: AC
authors.lastname: Yu
authors.forename: Xiao Qian
authors.initials: XQ
authors.lastname: Chen
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D005914:Globins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D016501:Neurites / Q000378:metabolism*; D000078787:Neuroglobin; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D051381:Rats
chemical_list: D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D005914:Globins; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse
references.citation: Mol Neurobiol. 2010 Jun;41(2-3):218-31
references.pmid: 20467836
references.citation: Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):9155-60
references.pmid: 22615373
references.citation: J Neurosci. 2010 Jul 7;30(27):9306-15
references.pmid: 20610765
references.citation: J Biol Chem. 2010 Jan 8;285(2):1358-70
references.pmid: 19861414
references.citation: Glia. 2005 Apr 15;50(2):182-6
references.pmid: 15657899
references.citation: Biochemistry. 2005 Mar 1;44(8):2943-8
references.pmid: 15723537
references.citation: J Cell Biol. 2008 Sep 22;182(6):1185-200
references.pmid: 18809727
references.citation: Stroke. 2010 Mar;41(3):557-9
references.pmid: 20075359
references.citation: J Biol Chem. 2007 Jun 1;282(22):16423-33
references.pmid: 17389603
references.citation: J Neurosci. 2012 Jan 4;32(1):264-74
references.pmid: 22219288
references.citation: Mol Neurobiol. 2013 Feb;47(1):197-208
references.pmid: 23180278
references.citation: J Biol Chem. 2011 Dec 9;286(49):42679-42689
references.pmid: 21965683
references.citation: Glia. 2003 Jun;42(4):315-24
references.pmid: 12730952
references.citation: J Neurochem. 2012 Jan;120(1):157-64
references.pmid: 21496024
references.citation: Antioxid Redox Signal. 2010 Sep 15;13(6):769-81
references.pmid: 20367258
references.citation: Nat Rev Neurosci. 2002 Sep;3(9):694-704
references.pmid: 12209118
references.citation: Mol Pharmacol. 2013 May;83(5):1109-19
references.pmid: 23478801
references.citation: Brain Res Rev. 2007 Feb;53(2):287-311
references.pmid: 17079020
references.citation: J Neurosci Res. 2005 Mar 15;79(6):798-808
references.pmid: 15672442
references.citation: Neurobiol Aging. 2005 Feb;26(2):275-8
references.pmid: 15582755
references.citation: Nature. 2000 Sep 28;407(6803):520-3
references.pmid: 11029004
references.citation: Apoptosis. 2010 Apr;15(4):401-11
references.pmid: 20091232
references.citation: Philos Trans R Soc Lond B Biol Sci. 2006 Sep 29;361(1473):1575-92
references.pmid: 16939976
references.citation: Biochem J. 2012 Apr 15;443(2):439-50
references.pmid: 22257123
references.citation: Cell Mol Life Sci. 2009 Sep;66(18):2975-84
references.pmid: 19504044
references.citation: Science. 2002 May 31;296(5573):1648-9
references.pmid: 12040181
references.citation: J Biol Chem. 2003 Sep 19;278(38):36505-12
references.pmid: 12860983
references.citation: Cell Mol Life Sci. 2008 Jan;65(1):161-76
references.pmid: 17975707
references.citation: FASEB J. 2012 Jan;26(1):81-92
references.pmid: 21940995
references.citation: J Neurotrauma. 2007 Feb;24(2):391-410
references.pmid: 17376002
references.citation: Biochem Biophys Res Commun. 2008 Mar 21;367(4):893-8
references.pmid: 18198128
references.citation: J Biol Chem. 2001 Aug 10;276(32):30106-10
references.pmid: 11429401