Metal-Chelating Plastic MALDI (pMALDI) Chips for the Enhancement of Phosphorylated-Peptide/Protein Signals

A disposable polymeric pMALDI array with a universal metal cation-chelatable surface for pretreatment/signal enhancement of phosphoproteins and/or phosphopeptides in complex samples was developed. Acrylic acid N-hydroxysuccinimide ester and methyl methacrylate monomers were copolymerized in thin lay...

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Bibliographic Details
Published in:Journal of proteome research 2007-09, Vol.6 (9), p.3842-3848
Main Authors: Ibáñez, Alfredo J, Muck, Alexander, Svatoš, Aleš
Format: Article
Language:English
Subjects:
Adsorption
Angiotensin II - chemistry
Gallium - chemistry
Humans
Mass Spectrometry - methods
Peptide Mapping
Peptides - chemistry
Phosphopeptides
Phosphoproteins - chemistry
Phosphorylation
Polymers - chemistry
Proteins - chemistry
Proteomics - instrumentation
Proteomics - methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Summary:A disposable polymeric pMALDI array with a universal metal cation-chelatable surface for pretreatment/signal enhancement of phosphoproteins and/or phosphopeptides in complex samples was developed. Acrylic acid N-hydroxysuccinimide ester and methyl methacrylate monomers were copolymerized in thin layer molds in a 1:13.3 molar ratio and subsequently treated with Nα,Nα-bis(carboxymethyl)-l-lysine to obtain a structured planar MALDI array. The prepared NTA pMALDI chip array was activated with metal cations (e.g., Ga(III), Ni(II)), and the selectivities for phosphopeptides (e.g., trypsin-digested α-casein (α-Cas), and phospho-angiotensin II (p-Ang)) were evaluated using MALDI-TOF/MS. The highest selectivity for proteins was observed for the Ni(II)−NTA chip. The p-Ang was enriched in the presence of BSA tryptic peptides ca. 5 times and represented the major peak after sample adsorption/washing on Ga(III)−NTA chip. The performance of the Ga(III)-chip, tested on α-Cas tryptic digest, is fully comparable to commercial systems. Additionally, higher MW peptides and limited methionine oxidation were observed with the chip. A combination of selective absorption of phosphoproteins on Ni(II)-chips and the further enrichment of digested phosphopeptides on the Ga(III)-chip can prove to be very useful for fast identification of unknown proteins using MALDI-TOF/MS. Keywords: phosphoproteomics • nickel(II) • gallium(III) • selective adsorption • phosphopeptide • phosphoprotein
ISSN:1535-3893
1535-3907
DOI:10.1021/pr070243r