Conformational chemistry of surface-attached calmodulin detected by acoustic shear wave propagation.
A thickness shear-mode acoustic wave device, operated in a flow-through format, was used to detect the binding of ions or peptides to surface-attached calmodulin. On-line surface attachment of the protein was achieved by immobilisation of the biotinylated molecule via a neutravidin-biotin linkage onto the surface of the gold electrode of the detector. The interaction between calmodulin, and calcium and magnesium ions induced an increase in resonant frequency and a decrease in motional resistance, which were reversible on washing with buffer. Interestingly, the changes in resonant frequency and motional resistance induced by the binding were opposite to the normal operation of the detector. The response was interpreted as a decrease in surface coupling (partial slip at the liquid/solid interface) instigated by exposure of hydrophobic domains on the protein, and an increase in the thickness, and hence effective wavelength, of the acoustic device, corresponding to an increase in the length of calmodulin by 1.5 A. This result is consistent with the literature value of 4 A. In addition, the interaction of the protein with peptide together with calcium ions was detected successfully, despite the relatively low molecular mass of the 2-kDa peptide. These results confirm the potential of acoustic wave physics for the detection of changes in the conformational chemistry of monolayer of biochemical macromolecules at the solid/liquid interface.
Wang
Xiaomeng
X
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario, CanadaM5S 3H6.
Ellis
Jonathan S
JS
Lyle
Emma-Louise
EL
Sundaram
Priyanka
P
Thompson
Michael
M
D000162:Acoustics; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D001360:Avidin / Q000378:metabolism; D019921:Biotinylation; D002021:Buffers; D002122:Calcium Chloride / Q000378:metabolism; D002147:Calmodulin / Q000032:analysis* / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013499:Surface Properties
D001059:Apoproteins; D002021:Buffers; D002147:Calmodulin; C093483:neutravidin; D001360:Avidin; D002122:Calcium Chloride