Detection and identification of plasma proteins that bind GlialCAM using ProteinChip arrays, SELDI-TOF MS, and nano-LC MS/MS
| Autor: | Edith Magnenat, Bruno Antonsson, Francis Vilbois, Ioannis Xenarios, Ursula Boschert, Pascale Sattonnet-Roche, Linda Favre-Kontula, Amanda E. I. Proudfoot |
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| Rok vydání: | 2008 |
| Předmět: |
Immunoprecipitation
Protein Array Analysis Cell Cycle Proteins Plasma protein binding Proteomics Biochemistry Mannose-Binding Lectin Protein–protein interaction Mice Affinity chromatography Peptide mass fingerprinting Tandem Mass Spectrometry Protein Interaction Mapping Animals Humans Nanotechnology Cell Cycle Protein Molecular Biology Chromatography Properdin Chemistry Cell Adhesion Molecules Neuron-Glia Interleukin-18 Proteins Blood proteins Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Intercellular Signaling Peptides and Proteins Chromatography Liquid Protein Binding |
| Zdroj: | Proteomics. 8(2) |
| ISSN: | 1615-9853 |
| Popis: | In order to fully understand biological processes it is essential to identify interactions in protein complexes. There are several techniques available to study this type of interactions, such as yeast two-hybrid screens, affinity chromatography, and coimmunoprecipitation. We propose a novel strategy to identify protein-protein interactions, comprised of first detecting the interactions using ProteinChips and SELDI-TOF MS, followed by the isolation of the interacting proteins through affinity beads and RP-HPLC and finally identifying the proteins using nano-LC MS/MS. The advantages of this new strategy are that the primary high-throughput screening of samples can be performed with small amounts of sample, no specific antibody is needed and the proteins represented on the SELDI-TOF MS spectra can be identified with high confidence. Furthermore, the method is faster and less labor-intensive than other current approaches. Using this novel method, we isolated and identified the interactions of two mouse plasma proteins, mannose binding lectin C and properdin, with GlialCAM, a type 1 transmembrane glycoprotein that belongs to the Ig superfamily. |
| Databáze: | OpenAIRE |
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