Solvent-induced conformational changes of polypeptides probed by electrospray-ionization mass spectrometry.
Electrospray-ionization (ESI) mass spectrometry is used to monitor higher order structural changes of polypeptides induced by alteration of the pH or organic solvent composition in the protein solution environment. A bimodal charge-state distribution is observed in the ESI mass spectrum of ubiquitin (relative molecular mass 8565) in solutions containing small amounts (less than 20%) of organic solvents. The distribution of peaks at high m/z (low-charge state) is found to represent the protein in its native, globular state; the higher-charge-state distribution is characteristic for a more extended conformation. Addition of methanol denaturant in excess of 40% v/v is needed to eliminate the low-charge-state distribution completely. Lesser amounts of acetonitrile, acetone, or isopropanol (approximately 20%) are required to denature the ubiquitin protein. Other proteins showing conformational effects in their ESI mass spectra are also illustrated. While the ESI spectra are related to solution phase structure, ESI-tandem mass spectrometry of multiply charged molecular ions of different conformation is suggested as a probe of gas-phase protein three-dimensional structure.
Loo
J A
JA
Chemical Sciences Department, Pacific Northwest Laboratory, Richland, WA 99532.
Loo
R R
RR
Udseth
H R
HR
Edmonds
C G
CG
Smith
R D
RD
D000818:Animals; D002417:Cattle; D002645:Chickens; D013058:Mass Spectrometry / Q000379:methods; D009113:Muramidase / Q000187:drug effects; D010455:Peptides / Q000187:drug effects*; D011487:Protein Conformation / Q000187:drug effects; D012997:Solvents / Q000494:pharmacology*; D014452:Ubiquitins / Q000187:drug effects
D010455:Peptides; D012997:Solvents; D014452:Ubiquitins; D009113:Muramidase