Preparation and characterization of bacterial protein complexes for structural analysis.
Bacteria mediate a large variety of biological processes using protein complexes. These complexes range from simple binary heterodimeric enzymes to more complex multi-subunit complexes that can be described as macromolecular machines. A key to understanding how these complexes function is obtaining structural information using methods that include electron microscopy, small-angle X-ray scattering, NMR spectroscopy, and X-ray crystallography. Here we describe a variety of approaches to the expression, purification, and biophysical characterization of bacterial protein complexes as a prerequisite to structural analysis. We also give several examples of the kinds of information these different biophysical approaches can provide and various experimental approaches to obtaining structure information for a given system. Further, we describe several examples of protein complexes where we have obtained structural data that have led to new biological insights.
Matte
Allan
A
Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, Canada.
Kozlov
Guennadi
G
Trempe
Jean-François
JF
Currie
Mark A
MA
Burk
David
D
Jia
Zongchao
Z
Gehring
Kalle
K
Ekiel
Irena
I
Berghuis
Albert M
AM
Cygler
Miroslaw
M
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*
D001426:Bacterial Proteins