Atomic force microscopy captures length phenotypes in single proteins.
We use single-protein atomic force microscopy techniques to detect length phenotypes in an Ig module. To gain amino acid resolution, we amplify the mechanical features of a single module by engineering polyproteins composed of up to 12 identical repeats. We show that on mechanical unfolding, mutant polyproteins containing five extra glycine residues added to the folded core of the module extend 20 A per module farther than the wild-type polyproteins. By contrast, similar insertions near the N or C termini have no effect. Hence, our atomic force microscopy measurements readily discriminate the location of the insert and measure its size with a resolution similar to that of NMR and x-ray crystallography.
Carrion-Vazquez
M
M
Department of Physiology, Mayo Foundation, Rochester, MN 55905, USA.
Marszalek
P E
PE
Oberhauser
A F
AF
Fernandez
J M
JM
D064211:Connectin; D018625:Microscopy, Atomic Force; D009124:Muscle Proteins / Q000737:chemistry*; D015202:Protein Engineering; D017510:Protein Folding; D011494:Protein Kinases / Q000737:chemistry*
D064211:Connectin; D009124:Muscle Proteins; D011494:Protein Kinases