Identifying Ca2+-binding sites in proteins by liquid chromatography-mass spectrometry using Ca2+-directed dissociations.

Azadeh Jamalian, Evert-Jan Sneekes, Hans Wienk, Lennard J M Dekker, Paul J A Ruttink, Mario Ursem, Theo M Luider, Peter C Burgers

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Article in Molecular & cellular proteomics : MCP, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

8 authors.

Azadeh JamalianFrom the ‡Department of Neurology, Laboratory of Neuro-Oncology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands; §Thermo Fisher Scientific/Dionex, Abberdaan 114, 1046 AA, Amsterdam, The Netherlands;

Evert-Jan SneekesFrom the ‡Department of Neurology, Laboratory of Neuro-Oncology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands; §Thermo Fisher Scientific/Dionex, Abberdaan 114, 1046 AA, Amsterdam, The Netherlands;

Hans Wienk¶NMR Spectroscopy, Bijvoet Center for Biomolecular Research, P.O. Box 80.075, 3508 TB, Utrecht University, Utrecht, The Netherlands;

Lennard J M DekkerFrom the ‡Department of Neurology, Laboratory of Neuro-Oncology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands;

Paul J A Ruttink‖Theoretical Chemistry Group, University of Utrecht, 3508 TC, Utrecht, The Netherlands.

Mario Ursem§Thermo Fisher Scientific/Dionex, Abberdaan 114, 1046 AA, Amsterdam, The Netherlands;

Theo M LuiderFrom the ‡Department of Neurology, Laboratory of Neuro-Oncology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands;

Peter C BurgersFrom the ‡Department of Neurology, Laboratory of Neuro-Oncology, Erasmus Medical Center, P.O. Box 2040, 3000 CA, Rotterdam, The Netherlands; p.burgers@erasmusmc.nl.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Here we describe a new method to identify calcium-binding sites in proteins using high-resolution liquid chromatography-mass spectrometry in concert with calcium-directed collision-induced dissociations. Our method does not require any modifications to the liquid chromatography-mass spectrometry apparatus, uses standard digestion protocols, and can be applied to existing high-resolution MS data files. In contrast to NMR, our method is applicable to very small amounts of complex protein mixtures (femtomole level). Calcium-bound peptides can be identified using three criteria: (1) the calculated exact mass of the calcium containing peptide; (2) specific dissociations of the calcium-containing peptide from threonine and serine residues; and (3) the very similar retention times of the calcium-containing peptide and the free peptide.

Indexed as

AnimalsBinding SitesCalciumCalcium-Binding ProteinsCell Cycle ProteinsChickensChromatography, High Pressure LiquidELAV ProteinsFemaleHumansPeptide MappingPlacentaPre-EclampsiaPregnancyS100 Calcium Binding Protein A6S100 ProteinsCalciumCalcium-Binding ProteinsCell Cycle ProteinsELAV ProteinsS100A6 protein, humanS100 Calcium Binding Protein A6S100 Proteins

Identifiers

PMID25023127

DOI10.1074/mcp.M114.038182

PMCPMC4223500

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.