title: Constant-time multidimensional electrophoretic NMR.
abstract: Multidimensional electrophoretic NMR (ENMR) has been introduced to determine structures of coexisting proteins and protein conformations in solution. Signals of different proteins are separated in a new dimension of electrophoretic flow according to their characteristic electrophoretic mobilities. The electrophoretic interferograms have been generated in the flow dimension in two approaches by incrementing either the amplitude or the duration of the electric field. The ENMR method of incrementing the duration of the electric field, however, introduces severe signal decays due to molecular diffusion and spin relaxation, limiting the effectiveness of the method. In this study, an improved method of constant-time multidimensional ENMR (CT-ENMR) has been proposed and successfully tested. The time delays between the magnetic field gradients and the RF pulses are kept constant in this new method so that the molecular diffusion and spin relaxation processes contribute to only a constant factor of signal amplitude. As an alternative approach of incrementing the amplitude of the electric field, this novel method significantly enhances our capability and potential in characterizing structural changes of interacting proteins during biological signaling processes. The CT-ENMR method is particularly useful in studies where the amplitude-incrementing of the electric field is not optimal. For example, the CT-ENMR method is superior when the electric field is applied in the direction not parallel to the static magnetic field B(0) to the xy-magnetization. The new method was successfully demonstrated with a sample solution containing 100 mM 4,9-dioxa-1,12-dodecanediamine and 100 mM L-aspartic acid in D(2)O.
authors.affiliation: Department of Medical Physics, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10021, USA.
authors.forename: Ercheng
authors.initials: E
authors.lastname: Li
authors.forename: Qiuhong
authors.initials: Q
authors.lastname: He