title: The ZIP5 ectodomain co-localizes with PrP and may acquire a PrP-like fold that assembles into a dimer.
abstract: The cellular prion protein (PrP(C)) was recently observed to co-purify with members of the LIV-1 subfamily of ZIP zinc transporters (LZTs), precipitating the surprising discovery that the prion gene family descended from an ancestral LZT gene. Here, we compared the subcellular distribution and biophysical characteristics of LZTs and their PrP-like ectodomains. When expressed in neuroblastoma cells, the ZIP5 member of the LZT subfamily was observed to be largely directed to the same subcellular locations as PrP(C) and both proteins were seen to be endocytosed through vesicles decorated with the Rab5 marker protein. When recombinantly expressed, the PrP-like domain of ZIP5 could be obtained with yields and levels of purity sufficient for structural analyses but it tended to aggregate, thereby precluding attempts to study its structure. These obstacles were overcome by moving to a mammalian cell expression system. The subsequent biophysical characterization of a homogeneous preparation of the ZIP5 PrP-like ectodomain shows that this protein acquires a dimeric, largely globular fold with an α-helical content similar to that of mammalian PrP(C). The use of a mammalian cell expression system also allowed for the expression and purification of stable preparations of Takifugu rubripes PrP-1, thereby overcoming a key hindrance to high-resolution work on a fish PrP(C).
authors.affiliation: Tanz Centre for Research in Neurodegenerative Diseases, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Cosmin L
authors.initials: CL
authors.lastname: Pocanschi
authors.forename: Sepehr
authors.initials: S
authors.lastname: Ehsani
authors.forename: Mohadeseh
authors.initials: M
authors.lastname: Mehrabian
authors.forename: Holger
authors.initials: H
authors.lastname: Wille
authors.forename: William
authors.initials: W
authors.lastname: Reginold
authors.forename: William S
authors.initials: WS
authors.lastname: Trimble
authors.forename: Hansen
authors.initials: H
authors.lastname: Wang
authors.forename: Adelinda
authors.initials: A
authors.lastname: Yee
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Bozóky
authors.forename: Lewis E
authors.initials: LE
authors.lastname: Kay
authors.forename: Julie D
authors.initials: JD
authors.lastname: Forman-Kay
authors.forename: James M
authors.initials: JM
authors.lastname: Rini
authors.forename: Gerold
authors.initials: G
authors.lastname: Schmitt-Ulms
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D027682:Cation Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018621:PrPC Proteins / Q000378:metabolism*; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism
chemical_list: D027682:Cation Transport Proteins; D018621:PrPC Proteins; C494728:Slc39a5 protein, mouse; D020696:rab5 GTP-Binding Proteins
references.citation: Bioinformatics. 2000 Jun;16(6):566-7
references.pmid: 10980157
references.citation: FEBS J. 2008 Jan;275(2):263-70
references.pmid: 18070106
references.citation: J Comput Chem. 2004 Oct;25(13):1605-12
references.pmid: 15264254
references.citation: PLoS One. 2011;6(10):e26800
references.pmid: 22046361
references.citation: J Neurosci. 2012 Sep 19;32(38):13255-63
references.pmid: 22993441
references.citation: Cell. 1986 Aug 1;46(3):417-28
references.pmid: 2873895
references.citation: Prion. 2011 Apr-Jun;5(2):88-92
references.pmid: 21628994
references.citation: Biochem J. 2009 Dec 23;425(2):341-51
references.pmid: 19888917
references.citation: Biometals. 2012 Apr;25(2):319-35
references.pmid: 22113231
references.citation: Biochim Biophys Acta. 2007 Feb;1769(2):117-23
references.pmid: 17292978
references.citation: Proc Natl Acad Sci U S A. 2013 Mar 26;110(13):5004-9
references.pmid: 23476064
references.citation: Hum Mol Genet. 2004 Mar 1;13(5):563-71
references.pmid: 14709598
references.citation: J Biol Chem. 2004 Dec 3;279(49):51433-41
references.pmid: 15322118
references.citation: Nucleic Acids Res. 2011 Jul;39(Web Server issue):W38-44
references.pmid: 21715387
references.citation: J Biol Chem. 2011 Nov 18;286(46):40255-65
references.pmid: 21917916
references.citation: PLoS Pathog. 2009 Oct;5(10):e1000608
references.pmid: 19798432
references.citation: FEBS Lett. 1997 Nov 17;417(3):400-4
references.pmid: 9409760
references.citation: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13363-83
references.pmid: 9811807
references.citation: PLoS One. 2009 Sep 28;4(9):e7208
references.pmid: 19784368
references.citation: Mol Cell Biol. 2009 Jan;29(1):129-39
references.pmid: 18936158
references.citation: FEBS J. 2009 Apr;276(8):2359-67
references.pmid: 19348007
references.citation: J Biomol NMR. 1995 Nov;6(3):277-93
references.pmid: 8520220
references.citation: Biochem Biophys Res Commun. 2000 Jan 7;267(1):134-8
references.pmid: 10623587
references.citation: Mol Med. 2007 Jul-Aug;13(7-8):396-406
references.pmid: 17673939
references.citation: Mol Cell Biol. 2003 Apr;23(7):2501-14
references.pmid: 12640132
references.citation: PLoS One. 2011;6(6):e21526
references.pmid: 21738690
references.citation: Mol Biol Rep. 2010 Jun;37(5):2295-300
references.pmid: 19685161
references.citation: J Biol Chem. 2004 Nov 19;279(47):49082-90
references.pmid: 15358787
references.citation: PLoS Biol. 2009 Mar 10;7(3):e55
references.pmid: 19278297
references.citation: Nat Biotechnol. 2003 Nov;21(11):1266-7
references.pmid: 14595348
references.citation: Annu Rev Nutr. 2009;29:153-76
references.pmid: 19400752
references.citation: Structure. 2013 Feb 5;21(2):236-46
references.pmid: 23290724
references.citation: Nature. 2004 May 20;429(6989):298-302
references.pmid: 15129296
references.citation: PLoS One. 2009 Jul 28;4(7):e6175
references.pmid: 19636413
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):651-5
references.pmid: 15647366
references.citation: J Cell Sci. 1999 Nov;112 ( Pt 21):3667-75
references.pmid: 10523503
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13419-24
references.pmid: 12370423
references.citation: J Mol Biol. 2012 Sep 28;422(4):556-574
references.pmid: 22687393
references.citation: Biochem J. 2008 Mar 1;410(2):237-44
references.pmid: 18254729
references.citation: PLoS One. 2011;6(9):e24624
references.pmid: 21931781
references.citation: Adv Protein Chem. 2001;57:55-82
references.pmid: 11447697
references.citation: Protein Sci. 1992 Oct;1(10):1343-52
references.pmid: 1363897
references.citation: J Biol Chem. 2002 Sep 6;277(36):33311-8
references.pmid: 12070160
references.citation: Biochemistry. 1999 Apr 27;38(17):5362-77
references.pmid: 10220323