title: Mass spectrometric analysis of mono- and multi-phosphopeptides by selective binding with NiZnFe₂O₄ magnetic nanoparticles.
abstract: Selective isolation of mono- and multi-phosphorylated peptides is important for understanding how a graded protein kinase or phosphatase signal can precisely modulate the on and off states of signal transduction pathways. Here we report that metal ions at exposed octahedral sites of nano-ferrites, including Fe3O4, NiFe₂O₄, ZnFe₂O₄ and NiZnFe₂O₄, have distinctly selective coordination abilities with mono- and multi- phosphopeptides. Due to their intrinsic magnetic properties and high surface area to volume ratios, these nanoparticles enable the rapid isolation of mono- and multi-phosphopeptides by an external magnetic field. Model phosphoprotein α-casein and two synthesized mono- and di-phosphopeptides have been chosen for proof-of-principle demonstrations, and these nanoparticles have also been applied to phosphoproteome profiling of zebrafish eggs. It is shown that NiZnFe₂O₄ is highly selective for multi-phosphopeptides. In contrast, Fe₃O₄, NiFe₂O₄ and ZnFe₂O₄ can bind with both mono- and multi-phosphopeptides with relatively stronger affinity towards mono-phosphopeptides.
authors.affiliation: Key Laboratory of Pesticides and Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, China. hyzhong@mail.ccnu.edu.cn
authors.forename: Hongying
authors.initials: H
authors.lastname: Zhong
authors.forename: Xiao
authors.initials: X
authors.lastname: Xiao
authors.forename: Shi
authors.initials: S
authors.lastname: Zheng
authors.forename: Wenyang
authors.initials: W
authors.lastname: Zhang
authors.forename: Mengjie
authors.initials: M
authors.lastname: Ding
authors.forename: Haiying
authors.initials: H
authors.lastname: Jiang
authors.forename: Lulu
authors.initials: L
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Kang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005290:Ferric Compounds / Q000737:chemistry*; D008280:Magnetics; D053768:Metal Nanoparticles; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry*; D010766:Phosphorylation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D015027:Zebrafish
chemical_list: D005290:Ferric Compounds; D010455:Peptides; D010750:Phosphoproteins
references.citation: Rapid Commun Mass Spectrom. 2010 May 30;24(10):1467-74
references.pmid: 20411586
references.citation: Nat Protoc. 2006;1(4):1929-35
references.pmid: 17487178
references.citation: PLoS Comput Biol. 2012;8(6):e1002551
references.pmid: 22737061
references.citation: Science. 2006 Aug 18;313(5789):976-9
references.pmid: 16917065
references.citation: J Proteome Res. 2012 May 4;11(5):2697-709
references.pmid: 22509746
references.citation: Nat Struct Mol Biol. 2012 Jan 15;19(2):171-5
references.pmid: 22245969
references.citation: Anal Chim Acta. 2012 Apr 29;724:73-9
references.pmid: 22483212
references.citation: Nat Rev Mol Cell Biol. 2008 Oct;9(10):815-20
references.pmid: 18784729
references.citation: Nature. 2011 Oct 12;480(7375):128-31
references.pmid: 21993622
references.citation: Proteomics. 2010 Feb;10(3):380-93
references.pmid: 19953538
references.citation: Annu Rev Biochem. 2009;78:797-825
references.pmid: 19489734
references.citation: Environ Health Perspect. 1995 Oct;103 Suppl 7:173-8
references.pmid: 8593867
references.citation: Mol Cell Proteomics. 2007 Dec;6(12):2088-99
references.pmid: 17698882
references.citation: Science. 1966 Jan 14;151(3707):172-7
references.pmid: 17746330
references.citation: Biochem Biophys Res Commun. 2010 Nov 19;402(3):537-42
references.pmid: 20971064
references.citation: Dev Biol. 2006 Jul 15;295(2):604-14
references.pmid: 16698010
references.citation: Eur J Biochem. 2001 Oct;268(19):5001-10
references.pmid: 11589691
references.citation: J Proteome Res. 2006 Sep;5(9):2431-7
references.pmid: 16944956
references.citation: Proc Natl Acad Sci U S A. 2010 Nov 9;107(45):19290-5
references.pmid: 20962272
references.citation: Nat Protoc. 2008;3(10):1630-8
references.pmid: 18833199
references.citation: Angew Chem Int Ed Engl. 2010 Mar 1;49(10):1862-6
references.pmid: 20155773
references.citation: Nat Rev Genet. 2007 May;8(5):353-67
references.pmid: 17440532
references.citation: Science. 1981 Apr 17;212(4492):298-304
references.pmid: 7209528
references.citation: Mol Cell Proteomics. 2008 Apr;7(4):661-71
references.pmid: 18039691
references.citation: Nat Rev Neurosci. 2011 May;12(5):248-9
references.pmid: 21505513
references.citation: Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):153-8
references.pmid: 14660795
references.citation: Trends Biochem Sci. 2000 Dec;25(12):596-601
references.pmid: 11116185
references.citation: Mol Cell Proteomics. 2007 Nov;6(11):2032-42
references.pmid: 17675664
references.citation: Mol Cell. 2006 Feb 17;21(4):481-93
references.pmid: 16483930
references.citation: Anal Chem. 2010 Sep 1;82(17):7249-58
references.pmid: 20712324
references.citation: Cell. 2006 Nov 3;127(3):635-48
references.pmid: 17081983