Article (Scientific journals)
Investigation of Structure-Stabilizing Elements in Proteins by Ion Mobility Mass Spectrometry and Collision-Induced Unfolding.
Grifnée, Elodie; Kune, Christopher; Delvaux, Cédric et al.
2024In Journal of the American Society for Mass Spectrometry
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Keywords :
3D structure; IM-MS; limited digestion; partially trypsin digested proteins; peptides; proteins
Abstract :
[en] A recently developed proteolytic reactor, designed for protein structural investigation, was coupled to ion mobility mass spectrometry to monitor collisional cross section (CCS) evolution of model proteins undergoing trypsin-mediated mono enzymatic digestion. As peptides are released during digestion, the CCS of the remaining protein structure may deviate from the classical 2/3 power of the CCS-mass relationship for spherical structures. The classical relationship between CCS and mass (CCS = A × M2/3) for spherical structures, assuming a globular shape in the gas phase, may deviate as stabilizing elements are lost during digestion. In addition, collision-induced unfolding (CIU) experiments on partially digested proteins provided insights into the CCS resilience in the gas phase to ion activation, potentially due to the presence of stabilizing elements. The study initially investigated a model peptide ModBea (3 kDa), assessing the impact of disulfide bridges on CCS resilience in both reduced and oxidized forms. Subsequently, β-lactoglobulin (2 disulfide bridges), calmodulin (Ca2+ coordination cation), and cytochrome c (heme) were selected to investigate the influence of common structuring elements on CCS resilience. CIU experiments probed the unfolding process, evaluating the effect of losing specific peptides on the energy landscapes of partially digested proteins. Comparisons of the TWCCSN2→He to trend curves describing the CCS/mass relationship revealed that proteins with structure-stabilizing elements consistently exhibit TWCCSN2→He and greater resilience toward CIU compared to proteins lacking these elements. The integration of online digestion, ion mobility, and CIU provides a valuable tool for identifying structuring elements in biopolymers in the gas phase.
Disciplines :
Chemistry
Author, co-author :
Grifnée, Elodie  ;  Université de Liège - ULiège > Département de pharmacie > Chimie médicale
Kune, Christopher  ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Delvaux, Cédric  ;  Université de Liège - ULiège > Molecular Systems (MolSys)
Tilmant, Thomas ;  Université de Liège - ULiège > Molecular Systems (MolSys)
Quinton, Loïc  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Matagne, André  ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Mazzucchelli, Gabriel  ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Far, Johann  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
De Pauw, Edwin  ;  Université de Liège - ULiège > Département de chimie (sciences)
Language :
English
Title :
Investigation of Structure-Stabilizing Elements in Proteins by Ion Mobility Mass Spectrometry and Collision-Induced Unfolding.
Publication date :
25 April 2024
Journal title :
Journal of the American Society for Mass Spectrometry
ISSN :
1044-0305
eISSN :
1879-1123
Publisher :
Springer, New-York, Us ny
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 April 2024

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