Detection and characterization of individual intermolecular bonds using optical tweezers.

Abstract:

:The development of scanning probe techniques has made it possible to examine protein-protein interactions at the level of individual molecular pairs. A calibrated optical tweezers, along with immunoglobulin G (IgG)-coated polystyrene microspheres, has been used to detect individual surface-linked Staphylococcus protein A (SpA) molecules and to characterize the strength of the noncovalent IgG-SpA bond. Microspheres containing, on average, less than one IgG per contact area were held in the optical trap while an SpA-coated substrate was scanned beneath them at a distance of approximately 50 nm. This geometry allows the trapped bead to make contact with the surface, from bond formation to rupture, and results in an enhancement of the force applied to a bond due to leverage supplied by the bead itself. Experiments yielded median single-bond rupture forces from 25 to 44 pN for IgG from four mammalian species, in general agreement with predictions based on free energies of association obtained from solution equilibrium constants.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Stout AL

doi

10.1016/S0006-3495(01)76263-7

subject

Has Abstract

pub_date

2001-06-01 00:00:00

pages

2976-86

issue

6

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(01)76263-7

journal_volume

80

pub_type

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