title: Two-state transition between molten globule and unfolded states of acetylcholinesterase as monitored by electron paramagnetic resonance spectroscopy.
abstract: Cys-231 of Torpedo californica acetylcholinesterase (EC 3.1.1.7) was selectively labeled with the mercury derivative of a stable nitroxyl radical. In 1.5 M guanidinium chloride, this conjugate exists in a molten globule state (MG), whereas in 5 M denaturant, it is in an unfolded state (U). The transition between the two states is reversible. In the MG, the label is highly immobilized, whereas in the U, it is almost freely rotating. The clearly distinct electron paramagnetic resonance (EPR) spectra of the two states permits the study of this transition. Upon elevating the guanidinium chloride concentration, a decrease in the EPR signal of the MG occurs concomitantly with an increase in the U signal, the total intensity of the EPR spectra remaining constant. This behavior is characteristic of a two-state transition. The thermodynamic characteristics of this transition (delta G0 and m), whether estimated directly from the EPR data or from both CD and fluorescence data analyzed by assuming a two-state scheme, are in good agreement.
authors.affiliation: Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: D I
authors.initials: DI
authors.lastname: Kreimer
authors.forename: R
authors.initials: R
authors.lastname: Szosenfogel
authors.forename: D
authors.initials: D
authors.lastname: Goldfarb
authors.forename: I
authors.initials: I
authors.lastname: Silman
authors.forename: L
authors.initials: L
authors.lastname: Weiner
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D003545:Cysteine; D004557:Electric Organ / Q000201:enzymology; D004578:Electron Spin Resonance Spectroscopy; D019791:Guanidine; D006146:Guanidines; D007700:Kinetics; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D014101:Torpedo
chemical_list: D006146:Guanidines; D000110:Acetylcholinesterase; D019791:Guanidine; D003545:Cysteine