Dissecting Streptavidin-Biotin Interaction with a Laminar Flow Chamber

Abstract

A laminar flow chamber was used to study single molecule interactions between biotinylated surfaces and streptavidin-coated spheres subjected to a hydrodynamic drag lower than a piconewton. Spheres were tracked with 20 ms and 40 nm resolution. They displayed multiple arrests lasting between a few tens of milliseconds and several minutes or more. Analysis of about 500,000 positions revealed that streptavidin-biotin interaction was multiphasic: transient bound states displayed a rupture frequency of 5.3 s(-1) and a rate of transition toward a more stable configuration of 1.3 s(-1). These parameters did not display any significant change when the force exerted on bonds varied between 3.5 and 11 pN. However, the apparent rate of streptavidin-biotin association exhibited about 10-fold decrease when the wall shear rate was increased from 7 to 22 s(-1), which supports the existence of an energy barrier opposing the formation of the transient binding state. It is concluded that a laminar flow chamber can yield new and useful information on the formation of molecular bonds, and especially on the structure of the external part of the energy landscape of ligand-receptor complexes.

Dissecting Individual Ligand–receptor Bonds With a Laminar Flow Chamber

“…Similar conclusions were obtained when streptavidin-coated spheres were made to interact with biotinylated surfaces [11]. Transient binding states were observed under conditions where the forces imposed on bonds varied between 3.5 and 11 pN.…”

“…Probably the laminar flow chamber is most effective in revealing the properties of the external part of the energy landscape [11], since it allows to probe bonds within a millisecond after formation, subjecting them to forces as low as a few piconewtons.…”

Dynamic force spectroscopy on multiple bonds: Experiments and model

“…as has been reported before for similar experimental setups [16,23]. The Poisson distribution contains the single parameter λ which determines the average number of broken bonds as…”

“…Experimentally, the streptavidin-biotin system is the best studied example for molecular bonding. In detail, it has been investigated with AFM [4], BFP [8] and flow chambers [16]. In the latter case, multiple bonds have been probed, but a persistent problem with flow chambers is that force is distributed in a heterogeneous way over the different bonds, thus rendering a quantitative analysis difficult.…”

Electromagnetophoretic Force Measurement of a Single Binding Interaction between Lectin and Yeast Cell Surfaces

“…A number of methods, such as hydrodynamic-flow-based method, 3 laser tweezers 4 and atomic force microscopy (AFM), 5 have been invented to investigate the interaction force between two different molecules, between a cell surface and a molecule, or between two different cells.…”