Comparative analysis of protein conformations by using 2D NOESY NMR spectra

摘要

The present disclosure provides a method for determining the relative conformations of a protein provided in different protein preparations, comprising steps of: (i) obtaining a first 2D NOESY NMR spectrum of a first protein preparation; (ii) obtaining a second 2D NOESY NMR spectrum of a second protein preparation; and (iii); determining whether a protein has a different conformation in the first and second protein preparations by comparing one or more cross-peaks in the first 2D NOESY NMR spectrum with one or more corresponding cross-peaks in the second 2D NOESY NMR spectrum.

主权项

1. A method of measuring differences in relative conformations of a recombinant glycoprotein provided in different preparations, comprising steps of:
generating a first 2D NOESY NMR spectrum of a first preparation of a recombinant glycoprotein using an NMR spectrometer, wherein the recombinant glycoprotein has been recombinantly expressed in a mammalian cell; generating a second 2D NOESY NMR spectrum, comparable to the first spectrum, of a second preparation of the same recombinant glycoprotein using an NMR spectrometer, wherein the recombinant gycoprotein has been recombinantly expressed in a mammalian cell; and measuring differences in conformation of the recombinant glycoprotein in the first and second glycoprotein preparations by comparing one or more cross-peaks in the first 2D NOESY NMR spectrum with one or more corresponding cross-peaks in the second 2D NOESY NMR spectrum.