On‐plate‐selective enrichment of glycopeptides using boronic acid‐modified gold nanoparticles for direct MALDI‐QIT‐TOF MS analysis

In this study, an on‐plate‐selective enrichment method is developed for fast and efficient glycopeptide investigation. Gold nanoparticles were first spotted and sintered on a stainless‐steel plate, then modified with 4‐mercaptophenylboronic acid to provide porous substrate with large specific surface and dual functions. These spots were used to selectively capture glycopeptides from peptide mixtures and the captured target peptides could be analyzed by MALDI‐MS simply by deposition of 2,5‐dihydroxybenzoic acid matrix. Horseradish peroxidase was employed as a standard glycoprotein to investigate the enrichment efficiency. In this way, the enrichment, washing and detection steps can all be fulfilled on a single MALDI target plate. The relatively small sample amount needed, low detection limit and rapid selective enrichment have made this on‐plate strategy promising for online enrichment of glycopeptides, which could be applied in high‐throughput proteome research.

[1]  J. Bergquist,et al.  Downsizing proteolytic digestion and analysis using dispenser‐aided sample handling and nanovial matrix‐assisted laser/desorption ionization‐target arrays , 2001, Proteomics.

[2]  X. Wang,et al.  Hydrothermal Syntheses of Gold Nanocrystals: From Icosahedral to Its Truncated Form , 2008 .

[3]  K. Gruber,et al.  High-throughput protein characterization using mass spectrometric immunoassay. , 2002, Analytical biochemistry.

[4]  M. Kostrzewa,et al.  Exploring the binding profiles of ConA, boronic acid and WGA by MALDI-TOF/TOF MS and magnetic particles. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[5]  Y. Kato,et al.  Rapid discovery and identification of a tissue-specific tumor biomarker from 39 human cancer cell lines using the SELDI ProteinChip platform. , 2003, Biochemical and biophysical research communications.

[6]  Younan Xia,et al.  Shape-controlled synthesis of metal nanostructures: the case of silver. , 2005, Chemistry.

[7]  A. Andreasen,et al.  Electrodesorption Kinetics and Molecular Interactions at Negatively Charged Self-Assembled Thiol Monolayers in Electrolyte Solutions , 2001 .

[8]  Paul A. C. Cloos,et al.  Post-Translational Modifications of Proteins: Implications for Aging, Antigen Recognition, and Autoimmunity , 2004, Biogerontology.

[9]  Jing-lan Wang,et al.  Specific capture of phosphopeptides on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry targets modified by magnetic affinity nanoparticles. , 2007, Rapid communications in mass spectrometry : RCM.

[10]  M. Tajiri,et al.  Hydrophilic affinity isolation and MALDI multiple-stage tandem mass spectrometry of glycopeptides for glycoproteomics. , 2004, Analytical chemistry.

[11]  Randall W. Nelson,et al.  Exploring the limit of detection in biomolecular interaction analysis mass spectrometry (BIA/MS): Detection of attomole amounts of native proteins present in complex biological mixtures , 2000 .

[12]  G. Wright,et al.  Proteinchip® surface enhanced laser desorption/ionization (SELDI) mass spectrometry: a novel protein biochip technology for detection of prostate cancer biomarkers in complex protein mixtures , 1999, Prostate Cancer and Prostatic Diseases.

[13]  Robert Podolsky,et al.  Assessing the utility of SELDI‐TOF and model averaging for serum proteomic biomarker discovery , 2006, Proteomics.

[14]  C. Fenselau,et al.  A targeted proteomics approach to the rapid identification of bacterial cell mixtures by matrix‐assisted laser desorption/ionization mass spectrometry , 2004, Proteomics.

[15]  F. Frantzen,et al.  Protein-boronic acid conjugates and their binding to low-molecular-mass cis-diols and glycated hemoglobin. , 1995, Journal of chromatography. B, Biomedical applications.

[16]  Randall W. Nelson,et al.  Peer Reviewed: Biomolecular Interaction Analysis Mass Spectrometry. , 2000 .

[17]  J. Hirabayashi,et al.  Lectin affinity capture, isotope-coded tagging and mass spectrometry to identify N-linked glycoproteins , 2003, Nature Biotechnology.

[18]  H. Girault,et al.  Specific on-plate enrichment of phosphorylated peptides for direct MALDI-TOF MS analysis. , 2007, Journal of proteome research.

[19]  Ralph G. Nuzzo,et al.  ADSORPTION OF BIFUNCTIONAL ORGANIC DISULFIDES ON GOLD SURFACES , 1983 .

[20]  A. Stensballe,et al.  Simplified Sample Preparation Method for Protein Identification by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry: In-Gel Digestion on the Probe Surface , 2001, Proteomics.

[21]  S. König.  Target coatings and desorption surfaces in biomolecular MALDI‐MS , 2008, Proteomics.

[22]  A. Deelder,et al.  Glycopeptide analysis by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry reveals novel features of horseradish peroxidase glycosylation. , 2004, Rapid communications in mass spectrometry : RCM.

[23]  D. Astruc,et al.  Gold nanoparticles: assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology. , 2004, Chemical reviews.

[24]  X. Zhang,et al.  On‐plate digestion of proteins using novel trypsin‐immobilized magnetic nanospheres for MALDI‐TOF‐MS analysis , 2007, Proteomics.

[25]  R. Nelson,et al.  Biomolecular interaction analysis mass spectrometry , 2006 .

[26]  S. Hagiwara,et al.  Lactoferrin concentrations in milk from normal and subclinical mastitic cows. , 2003, The Journal of veterinary medical science.

[27]  Ruedi Aebersold,et al.  Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry , 2003, Nature Biotechnology.

[28]  Andreas Quandt,et al.  Annotated regions of significance of SELDI‐TOF‐MS spectra for detecting protein biomarkers , 2006, Proteomics.

[29]  R. Knochenmuss.  Ion formation mechanisms in UV-MALDI. , 2006, The Analyst.

[30]  Jakob Bunkenborg,et al.  A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation. , 2004, Journal of proteome research.

[31]  J. Roth.  Protein N-glycosylation along the secretory pathway: relationship to organelle topography and function, protein quality control, and cell interactions. , 2002, Chemical reviews.

[32]  T. Yamori,et al.  A novel method for analyzing phosphoproteins using SELDI‐TOF MS in combination with a series of recombinant proteins , 2007, Proteomics.

[33]  Yu-Chie Chen,et al.  Rapid enrichment of phosphopeptides from tryptic digests of proteins using iron oxide nanocomposites of magnetic particles coated with zirconia as the concentrating probes. , 2007, Journal of proteome research.

[34]  R. Dwek,et al.  Glycosylation and the immune system. , 2001, Science.

[35]  Eun Kyu Lee,et al.  Immobilization of aminophenylboronic acid on magnetic beads for the direct determination of glycoproteins by matrix assisted laser desorption ionization mass spectrometry , 2005, Journal of the American Society for Mass Spectrometry.

[36]  G. Lienhard,et al.  The binding of boronic acids to chymotrypsin. , 1974, Biochemistry.

[37]  N. O'donnell,et al.  Intracellular glycosylation and development. , 2002, Biochimica et biophysica acta.

[38]  Younan Xia,et al.  One‐Dimensional Nanostructures: Synthesis, Characterization, and Applications , 2003 .

[39]  A. Helenius,et al.  Intracellular functions of N-linked glycans. , 2001, Science.

[40]  Pengyuan Yang,et al.  Facile synthesis of aminophenylboronic acid-functionalized magnetic nanoparticles for selective separation of glycopeptides and glycoproteins. , 2008, Chemical communications.

[41]  Carolyn R. Bertozzi,et al.  Chemical Glycobiology , 2001, Science.