Details

Autor(en) / Beteiligte

• El-Baba, Tarick J
• Woodall, Daniel W
• Raab, Shannon A
• Fuller, Daniel R
• Laganowsky, Arthur
• Russell, David H
• Clemmer, David E

Titel

Melting Proteins: Evidence for Multiple Stable Structures upon Thermal Denaturation of Native Ubiquitin from Ion Mobility Spectrometry-Mass Spectrometry Measurements

Ist Teil von

• Journal of the American Chemical Society, 2017-05, Vol.139 (18), p.6306-6309

Ort / Verlag

United States: American Chemical Society

Erscheinungsjahr

2017

Quelle

MEDLINE

Beschreibungen/Notizen

• Ion mobility and mass spectrometry techniques are coupled with a temperature-controlled electrospray ionization source to follow the structural transitions of ubiquitin in aqueous solution (pH = 3) at elevated solution temperatures (T = 26–96 °C). Changes in the charge state distribution are consistent with a two-state, cooperative unfolding transition having a melting temperature of T m = 71 ± 2 °C, in agreement with prior measurements [ Wintrode, P. L. ; Makhatadze, G. I. ; Privalov, P. L. Proteins, 1994, 18, 246−253 ]. However, analysis of ion mobility distributions reveals the two-state transition is a composite of transitions involving at least nine unique species: three native or native-like structures; two that appear to be equilibrium intermediates (i.e., populations of new conformers that form at elevated temperatures but subsequently disappear at higher temperatures); and four products observed at high temperatures, including the well-characterized ubiquitin A state, and two solution species that are differentiated based on a cis- or trans-configured Glu18-Pro19 peptide bond. These nine states vary in abundances by factors as large as ∼103 over the range of solution temperatures. Although experimental melting transitions are conceived as a loss of well-defined structure leading to a random distribution of unstructured, denatured forms, the results provide evidence for new conformers having at least some well-defined structural elements are stabilized as temperature is increased.