Solution and gas-phase modifiers effect on heme proteins environment and conformational space

Abstract

Abstract The molecular environment is known to impact the secondary and tertiary structure of biomolecules, shifting the equilibrium between different conformational and oligomerization states. In the present study, the effect of solution additives and gas-phase modifiers on the molecular environment of two common heme proteins, bovine cytochrome c and equine myoglobin, is investigated as a function of the time after desolvation (e.g., 100 - 500 ms) using trapped ion mobility spectrometry – mass spectrometry. Changes in the mobility profiles are observed depending on the starting solution composition (i.e., in aqueous solution at neutral pH or in the presence of organic content: methanol, acetone, or acetonitrile) depending on the protein. In the presence of gas-phase modifiers (i.e., N 2 containing methanol, acetone, or acetonitrile), a shift in the mobility profiles driven by the gas-modifier mass and size and changes in the relative abundances and number of IMS bands are observed. We attribute these changes in the mobility profiles in the presence of gas-phase modifiers to a clustering/declustering mechanism by which organic molecules adsorb to the protein ion surface and lower energetic barriers for interconversion between conformational states, thus redefining the free energy landscape and equilibria between conformers. These structural biology experiments open new avenues for manipulation and interrogation of biomolecules in the gas-phase with the potential to emulate a large suite of solution conditions, ultimately including conditions that more accurately reflect a variety of intracellular environments.

Related Papers

• → Blue Myoglobin Reconstituted with an Iron Porphycene Shows Extremely High Oxygen Affinity(2002)123 cited
• → Life without myoglobin(2000)38 cited
• → Heme orientational disorder in reconstituted and native sperm whale myoglobin(1983)163 cited
• → NMR study of haem exchange reaction of native myoglobin and haemoglobin(1993)1 cited
• → Conversion of Hemoprotein Function by Chemical Modification(2002)