Residue-specific structural kinetics of proteins through the union of isotope labeling, mid-IR pulse shaping, and coherent 2D IR spectroscopy

TitleResidue-specific structural kinetics of proteins through the union of isotope labeling, mid-IR pulse shaping, and coherent 2D IR spectroscopy
Publication TypeJournal Article
Year of Publication2010
AuthorsMiddleton CT, Woys AM, Mukherjee SS, Zanni MT
Secondary TitleMethods
Volume52
Issue1
Pagination12 - 22
Date Published9/2010
Abstract

We describe a methodology for studying protein kinetics using a rapid-scan technology for collecting 2D IR spectra. In conjunction with isotope labeling, 2D IR spectroscopy is able to probe the secondary structure and environment of individual residues in polypeptides and proteins. It is particularly useful for membrane and aggregate proteins. Our rapid-scan technology relies on a mid-IR pulse shaper that computer generates the pulse shapes, much like in an NMR spectrometer. With this device, data collection is faster, easier, and more accurate. We describe our 2D IR spectrometer, as well as protocols for 13C18O isotope labeling, and then illustrate the technique with an application to the aggregation of the human islet amyloid polypeptide implicated in type 2 diabetes.

URLhttp://www.ncbi.nlm.nih.gov/pubmed/20472067
DOI10.1016/j.ymeth.2010.05.002.
Short TitleMethods