Biophysical assays for protein interactions in the Wsp sensory system and biofilm formation.
Many signal transduction and regulatory events are mediated by a change in oligomeric state upon posttranslational modification or ligand binding. Hence, the characterization of proteins and protein complexes with respect to their size and shape is crucial for elucidating the molecular mechanisms that control their activities. Commonly used methods for the determination of molecular weights of biological polymers such as standard size-exclusion chromatography or analytical ultracentrifugation have been applied successfully but have some limitations. Static multiangle light scattering presents an attractive alternative approach for absolute molecular weight measurements in solution. We review the biophysical principles, advantages, and pitfalls of some popular methods for determining the quaternary structure of proteins, using the response regulator diguanylate cyclase WspR from Pseudomonas and FimX, a protein involved in Pseudomonas aeruginosa twitching motility, as examples.
De
Nabanita
N
Department of Molecular Medicine, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
Navarro
Marcos V A S
MV
Wang
Qi
Q
Krasteva
Petya V
PV
Sondermann
Holger
H
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms / Q000254:growth & development*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism; D019761:Phosphorus-Oxygen Lyases / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D046912:Multiprotein Complexes; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase