Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry.

Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. Research Abstract Details 

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Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. Abstract Text:

Christopher T Veros,Neil J Oldham,Christopher T Veros,Neil J Oldham,

Affinity constants for the binding of a range of substrate and non-substrate oligosaccharides to hen egg white lysozyme were determined by direct observation of the protein.ligand complexes using electrospray ionisation mass spectrometry (ESI-MS) with a chip-based nano-ESI source. The values obtained for a series of beta-1,4-N-acetylglucosamine oligomers (NAGn) were found to be in good agreement with those determined by fluorescence measurement. Oligomers of alpha-1,4-glucose (Glcn), which are believed to bind to lysozyme non-specifically, exhibited a 10(6)- to 10(8)-fold lower affinity for the enzyme. Lysozyme.NAGn complexes displayed an increase in Ka from n=2 to n=4, but then reached a plateau. In contrast non-specific lysozyme.Glcn complexes showed no such trend. Determination of gas-phase complex stability was achieved by quantitative collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) measurements. The collision energy (Ec50) or laser power (IRMPD50) required to dissociate precursor ions to 50% of their original intensity was determined for lysozyme.NAGn and Glcn complexes using the [M+8H]8+ charge state. An excellent correlation between trends in Ka and gas-phase stability was seen for NAGn oligomers bound to lysozyme, whereas no such relationship was observed with the non-specific, weaker lysozyme.Glcn complexes. These results illustrate that ESI-MS can be used to quantify the interactions between lysozyme and oligosaccharides in both the solution and gas phase and that measurement of gas-phase complex stability by CID or IRMPD can provide information about specific solution binding events.

Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. Publishing Authors By Initials

CT Veros,NJ Oldham,CT Veros,NJ Oldham,

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Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Rapid communications in mass spectrometry : RCM

VOLUME: 21

Page Numbers: 3505-10

Journal Abbreviation: Rapid Commun. Mass Spectrom.

ISSN: 0951-4198

DAY: 7

MONTH: 11

YEAR: 2007

Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. Information

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LANGUAGE: eng

NlmUniqueID: 8802365

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Grant and Affiliation Information for Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry.

AFFILIATION: Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford OX1 3TA, UK.

Country: England

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MEDLINETA: Rapid Commun Mass Spectrom

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