title: Phosphopeptide enrichment with TiO2-modified membranes and investigation of tau protein phosphorylation.
abstract: Selective enrichment of phosphopeptides prior to their analysis by mass spectrometry (MS) is vital for identifying protein phosphorylation sites involved in cellular regulation. This study describes modification of porous nylon substrates with TiO2 nanoparticles to create membranes that rapidly enrich phosphopeptides. Membranes with a 22-mm diameter bind 540 nmol of phosphoangiotensin and recover 70% of the phosphopeptides in mixtures with a 15-fold excess of nonphosphorylated proteins. Recovery is 90% for a pure phosphopeptide. Insertion of small membrane disks into HPLC fittings allows rapid enrichment from 5 mL of 1 fmol/μL phosphoprotein digests and concentration into small-volume (tens of microliters) eluates. The combination of membrane enrichment with tandem mass spectrometry reveals seven phosphorylation sites from in vivo phosphorylated tau (p-tau) protein, which is associated with Alzheimer's disease.
authors: Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
authors: Yu-Jing
authors: YJ
authors: Tan
authors: Dexin
authors: D
authors: Sui
authors: Wei-Han
authors: WH
authors: Wang
authors: Min-Hao
authors: MH
authors: Kuo
authors: Gavin E
authors: GE
authors: Reid
authors: Merlin L
authors: ML
authors: Bruening
mesh_terms: D006801:Humans; D008567:Membranes, Artificial; D010748:Phosphopeptides / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation / Q000502:physiology; D014025:Titanium / Q000737:chemistry / Q000378:metabolism*; D016875:tau Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D008567:Membranes, Artificial; D010748:Phosphopeptides; D016875:tau Proteins; C009495:titanium dioxide; D014025:Titanium
references: Nat Methods. 2007 Oct;4(10):798-806
references: 17901869
references: Nat Protoc. 2006;1(4):1929-35
references: 17487178
references: Proteomics. 2011 Jun;11(12):2568-72
references: 21598390
references: Nat Rev Drug Discov. 2007 Jun;6(6):464-79
references: 17541419
references: J Cell Sci. 1996 Jun;109 ( Pt 6):1537-43
references: 8799840
references: Mol Cell Proteomics. 2011 Oct;10(10):M110.006452
references: 21715320
references: J Proteome Res. 2010 Jan;9(1):430-43
references: 19899826
references: J Sep Sci. 2007 May;30(8):1207-13
references: 17595956
references: Nat Rev Mol Cell Biol. 2001 Oct;2(10):769-76
references: 11584304
references: Anal Chem. 2011 Oct 15;83(20):7635-44
references: 21899308
references: Nano Lett. 2006 Oct;6(10):2268-72
references: 17034095
references: Mass Spectrom Rev. 2011 Jul-Aug;30(4):600-25
references: 21294150
references: J Chromatogr A. 2008 May 23;1192(1):95-102
references: 18384799
references: Neuroscience. 2002;115(3):829-37
references: 12435421
references: J Chromatogr A. 2007 Sep 21;1165(1-2):128-35
references: 17714720
references: Int J Alzheimers Dis. 2011;2011:352805
references: 21776376
references: Anal Chem. 2011 Mar 1;83(5):1624-31
references: 21306131
references: J Proteome Res. 2008 Feb;7(2):687-97
references: 18034456
references: Proteomics. 2007 Feb;7(3):351-60
references: 17177250
references: J Neurochem. 2000 Apr;74(4):1587-95
references: 10737616
references: Anal Chem. 2010 Dec 15;82(24):10045-51
references: 21087034
references: Nature. 2003 Mar 13;422(6928):198-207
references: 12634793
references: Trends Mol Med. 2009 Mar;15(3):112-9
references: 19246243
references: Anal Chem. 2010 Apr 1;82(7):2907-15
references: 20199055
references: Proc Natl Acad Sci U S A. 2011 May 24;108(21):8577-82
references: 21606340
references: J Proteomics. 2011 Nov 18;74(12):2786-97
references: 21839867
references: J Proteome Res. 2008 Sep;7(9):4058-69
references: 18707149
references: Analyst. 2008 Sep;133(9):1128-38
references: 18709185
references: J Proteome Res. 2008 Dec;7(12):5167-76
references: 19367720
references: Biochim Biophys Acta. 2005 Jan 3;1739(2-3):179-97
references: 15615637
references: Nat Methods. 2007 Mar;4(3):231-7
references: 17293869
references: Mol Cell Proteomics. 2008 Apr;7(4):661-71
references: 18039691
references: J Biol Chem. 1997 Oct 3;272(40):25326-32
references: 9312151
references: Anal Biochem. 1986 Apr;154(1):250-4
references: 3085541
references: Anal Chem. 2008 Aug 1;80(15):5727-35
references: 18578546
references: Anal Chem. 2008 Dec 15;80(24):9735-47
references: 19012417
references: Anal Chem. 2011 Feb 1;83(3):761-6
references: 21210638
references: Cell. 2000 Jan 7;100(1):113-27
references: 10647936
references: J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Apr 1;891-892:109-12
references: 22406350
references: J Proteome Res. 2008 Feb;7(2):749-55
references: 18171022
references: J Proteome Res. 2010 Jun 4;9(6):3005-15
references: 20380454
references: J Proteome Res. 2008 Jun;7(6):2526-38
references: 18473453
references: Trends Biochem Sci. 2000 Dec;25(12):596-601
references: 11116185
references: Mol Biosyst. 2011 May;7(5):1420-9
references: 21327254
references: J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 15;878(5-6):515-24
references: 20075017
references: Cell. 2006 Nov 3;127(3):635-48
references: 17081983
references: Pharmacol Ther. 1999 May-Jun;82(2-3):111-21
references: 10454190
references: FEBS Lett. 1998 Sep 25;436(1):28-34
references: 9771888
references: Anal Chem. 2007 Jun 15;79(12):4666-73
references: 17523591
references: Anal Chem. 2004 Jul 15;76(14):3935-43
references: 15253627
references: Anal Chem. 2009 Jul 15;81(14):5794-805
references: 19522514
references: Proteomics. 2009 Mar;9(6):1451-68
references: 19235172
references: Mol Cell Proteomics. 2005 Jul;4(7):873-86
references: 15858219
references: J Proteome Res. 2008 Aug;7(8):3304-13
references: 18578522
references: J Proteome Res. 2008 May;7(5):1809-18
references: 18327898
references: Proteomics. 2005 Nov;5(16):4052-61
references: 16196093
references: J Chromatogr A. 2006 May 26;1116(1-2):38-45
references: 16580007
references: Mol Neurodegener. 2007 Jun 28;2:12
references: 17598919
references: Mass Spectrom Rev. 2010 Jan-Feb;29(1):29-54
references: 19263479