Collision induced unfolding of protein ions in the gas phase studied by ion mobility-mass spectrometry: the effect of ligand binding on conformational stability.
Ion mobility spectrometry, with subsequent mass spectrometric detection, has been employed to study the stability of compact protein conformations of FK-binding protein, hen egg-white lysozyme, and horse heart myoglobin in the presence and absence of bound ligands. Protein ions, generated by electrospray ionization from ammonium acetate buffer, were activated by collision with argon gas to induce unfolding of their compact structures. The collisional cross sections (Omega) of folded and unfolded conformations were measured in the T-Wave mobility cell of a Waters Synapt HDMS (Waters, Altrincham, UK) employing a calibration against literature values for a range of protein standards. In the absence of activation, collisional cross section measurements were found to be consistent with those predicted for folded protein structures. Under conditions of defined collisional activation energies partially unfolded conformations were produced. The degree of unfolding and dissociation induced by these defined collision energies are related to the stability of noncovalent intra- and intermolecular interactions within protein complexes. These findings highlight the additional conformational stability of protein ions in the gas phase resulting from ligand binding.
Hopper
Jonathan T S
JT
School of Chemistry, University of Nottingham, University Park, Nottingham, United Kingdom.
Oldham
Neil J
NJ
D000818:Animals; D002645:Chickens; D005740:Gases / Q000737:chemistry; D006736:Horses; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D055550:Protein Stability; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D013816:Thermodynamics
D005740:Gases; D009211:Myoglobin; D011506:Proteins; C421864:hen egg lysozyme; D009113:Muramidase