title: The Amyloid Precursor Protein of Alzheimer's Disease Clusters at the Organelle/Microtubule Interface on Organelles that Bind Microtubules in an ATP Dependent Manner.
abstract: The amyloid precursor protein (APP) is a causal agent in the pathogenesis of Alzheimer's disease and is a transmembrane protein that associates with membrane-limited organelles. APP has been shown to co-purify through immunoprecipitation with a kinesin light chain suggesting that APP may act as a trailer hitch linking kinesin to its intercellular cargo, however this hypothesis has been challenged. Previously, we identified an mRNA transcript that encodes a squid homolog of human APP770. The human and squid isoforms share 60% sequence identity and 76% sequence similarity within the cytoplasmic domain and share 15 of the final 19 amino acids at the C-terminus establishing this highly conserved domain as a functionally import segment of the APP molecule. Here, we study the distribution of squid APP in extruded axoplasm as well as in a well-characterized reconstituted organelle/microtubule preparation from the squid giant axon in which organelles bind microtubules and move towards the microtubule plus-ends. We find that APP associates with microtubules by confocal microscopy and co-purifies with KI-washed axoplasmic organelles by sucrose density gradient fractionation. By electron microscopy, APP clusters at a single focal point on the surfaces of organelles and localizes to the organelle/microtubule interface. In addition, the association of APP-organelles with microtubules is an ATP dependent process suggesting that the APP-organelles contain a microtubule-based motor protein. Although a direct kinesin/APP association remains controversial, the distribution of APP at the organelle/microtubule interface strongly suggests that APP-organelles have an orientation and that APP like the Alzheimer's protein tau has a microtubule-based function.
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: James W
authors.initials: JW
authors.lastname: Stevenson
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Eliza A
authors.initials: EA
authors.lastname: Conaty
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Rylie B
authors.initials: RB
authors.lastname: Walsh
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Poidomani
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Colin M
authors.initials: CM
authors.lastname: Samoriski
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Brianne J
authors.initials: BJ
authors.lastname: Scollins
authors.affiliation: Biology Department, Providence College, Providence, Rhode Island, United States of America.|Bell Center, Marine Biological Laboratory, Woods Hole, Massachusetts, United States of America.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: DeGiorgis
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000544:Alzheimer Disease / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D001369:Axons / Q000378:metabolism; D049832:Decapodiformes / Q000378:metabolism*; D006801:Humans; D008870:Microtubules / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism
chemical_list: D016564:Amyloid beta-Protein Precursor; D014404:Tubulin; D000255:Adenosine Triphosphate
references.citation: Neuron. 2001 Nov 8;32(3):389-401
references.pmid: 11709151
references.citation: J Cell Sci. 1998 Nov;111 ( Pt 21):3221-34
references.pmid: 9763516
references.citation: Neuropharmacology. 1999 Mar;38(3):349-59
references.pmid: 10219973
references.citation: J Cell Biol. 1987 Sep;105(3):1273-82
references.pmid: 2958482
references.citation: J Neurosci. 2005 Mar 2;25(9):2386-95
references.pmid: 15745965
references.citation: Neuron. 2000 Nov;28(2):449-59
references.pmid: 11144355
references.citation: Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10758-62
references.pmid: 1279693
references.citation: Elife. 2015 Nov 09;4:e09743
references.pmid: 26551565
references.citation: J Biol Chem. 2001 Jul 27;276(30):28014-21
references.pmid: 11382767
references.citation: J Neurosci. 2000 Nov 1;20(21):7951-63
references.pmid: 11050115
references.citation: Methods Mol Biol. 2001;164:1-7
references.pmid: 11217600
references.citation: PLoS One. 2013 Dec 04;8(12):e81926
references.pmid: 24324731
references.citation: Nature. 1991 Oct 31;353(6347):844-6
references.pmid: 1944558
references.citation: EMBO J. 1988 Apr;7(4):949-57
references.pmid: 2900137
references.citation: Biochem Biophys Res Commun. 1997 Nov 7;240(1):1-7
references.pmid: 9367871
references.citation: Exp Brain Res. 2012 Apr;217(3-4):353-64
references.pmid: 21960299
references.citation: Nature. 1991 Feb 21;349(6311):704-6
references.pmid: 1671712
references.citation: Curr Alzheimer Res. 2012 Feb;9(2):165-77
references.pmid: 21605033
references.citation: Development. 2004 May;131(9):2173-81
references.pmid: 15073156
references.citation: J Neurosci Res. 1991 Jan;28(1):90-100
references.pmid: 1645774
references.citation: Nature. 1987 Feb 19-25;325(6106):733-6
references.pmid: 2881207
references.citation: PLoS One. 2011 Apr 27;6(4):e18754
references.pmid: 21556148
references.citation: Traffic. 2008 Nov;9(11):1867-77
references.pmid: 18928504
references.citation: Cell. 1995 May 19;81(4):525-31
references.pmid: 7758106
references.citation: BMC Res Notes. 2011 Nov 01;4:476
references.pmid: 22044463
references.citation: J Cell Sci. 1996 May;109 ( Pt 5):999-1008
references.pmid: 8743947
references.citation: Neurosci Lett. 1995 Jun 9;192(2):132-6
references.pmid: 7675321
references.citation: J Biol Chem. 2004 Apr 30;279(18):18146-56
references.pmid: 14970212
references.citation: Cytoskeleton (Hoboken). 2011 Oct;68(10):566-77
references.pmid: 21913340
references.citation: Neurobiol Aging. 1997 Nov-Dec;18(6):661-9
references.pmid: 9461064
references.citation: Alzheimers Dement. 2007 Jul;3(3):186-91
references.pmid: 19595937
references.citation: BMC Bioinformatics. 2008 Jun 09;9:271
references.pmid: 18541026
references.citation: Proc Natl Acad Sci U S A. 1985 Jun;82(12):4245-9
references.pmid: 3159021
references.citation: Curr Biol. 2010 May 11;20(9):R413-5
references.pmid: 20462485
references.citation: Eur J Neurosci. 1995 Mar 1;7(3):501-10
references.pmid: 7773447
references.citation: Biochem Biophys Res Commun. 1984 May 16;120(3):885-90
references.pmid: 6375662
references.citation: Hum Mol Genet. 2013 Oct 1;22(19):3828-43
references.pmid: 23710041
references.citation: Cell. 1985 Dec;43(3 Pt 2):623-32
references.pmid: 2416467
references.citation: Cell. 1985 Feb;40(2):455-62
references.pmid: 2578325
references.citation: Proc Natl Acad Sci U S A. 1989 Mar;86(5):1548-52
references.pmid: 2466291
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16532-7
references.pmid: 17062754
references.citation: Proc Natl Acad Sci U S A. 1998 May 26;95(11):6460-4
references.pmid: 9600988
references.citation: Hum Mol Genet. 2014 Mar 15;23(6):1631-43
references.pmid: 24203698
references.citation: J Biol Chem. 2003 May 9;278(19):17401-7
references.pmid: 12611883
references.citation: Brain Res. 1992 Oct 2;592(1-2):353-7
references.pmid: 1280522
references.citation: Physiol Rev. 2001 Apr;81(2):741-66
references.pmid: 11274343
references.citation: Eur J Biochem. 1997 Dec 1;250(2):354-63
references.pmid: 9428684
references.citation: Science. 1982 Dec 10;218(4577):1129-31
references.pmid: 6183745
references.citation: J Cell Biol. 1982 Jul;94(1):1-6
references.pmid: 6889603
references.citation: J Cell Biol. 2003 Jan 6;160(1):113-23
references.pmid: 12515826
references.citation: J Cell Biol. 1992 Oct;119(2):389-99
references.pmid: 1400582
references.citation: Ann N Y Acad Sci. 2005 Jun;1048:149-65
references.pmid: 16154929
references.citation: Nature. 1992 Jun 11;357(6378):500-3
references.pmid: 1608449
references.citation: J Neurosci. 2009 Mar 18;29(11):3565-78
references.pmid: 19295161
references.citation: J Cell Sci. 1996 May;109 ( Pt 5):991-8
references.pmid: 8743946
references.citation: Science. 1982 Dec 10;218(4577):1127-9
references.pmid: 6183744
references.citation: Nature. 1966 Jan 1;209(5018):109-10
references.pmid: 5927229
references.citation: Biochemistry. 2000 Sep 5;39(35):10831-9
references.pmid: 10978169
references.citation: J Cell Biol. 2013 Aug 5;202(3):495-508
references.pmid: 23897889
references.citation: Cell. 1985 Aug;42(1):39-50
references.pmid: 3926325
references.citation: Cell. 1985 Feb;40(2):449-54
references.pmid: 2578324
references.citation: J Neurosci. 2009 Sep 2;29(35):10788-801
references.pmid: 19726636
references.citation: J Vis Exp. 2012 Apr 27;(62):
references.pmid: 22565641
references.citation: Nature. 1963 Jan 12;197:192-3
references.pmid: 14032480
references.citation: Nature. 1997 Jun 5;387(6633):569-72
references.pmid: 9177342
references.citation: Proc Natl Acad Sci U S A. 1990 Feb;87(4):1561-5
references.pmid: 1689489
references.citation: Neurosci Lett. 1991 Nov 25;133(1):1-2
references.pmid: 1791986
references.citation: Nat Genet. 1993 Sep;5(1):95-100
references.pmid: 8220435