Diffusing colloidal probes of protein and synthetic macromolecule interactions.
A new approach is described for measuring kT and nanometer scale protein-protein and protein-synthetic macromolecule interactions. The utility of this method is demonstrated by measuring interactions of bovine serum albumin (BSA) and copolymers with exposed polyethyleneoxide (PEO) moieties adsorbed to hydrophobically modified colloids and surfaces. Total internal reflection and video microscopy are used to track three-dimensional trajectories of many single diffusing colloids that are analyzed to yield interaction potentials, mean-square displacements, and colloid-surface association lifetimes. A criterion is developed to identify colloids as being levitated, associated, or deposited based on energetic, spatial, statistical, and temporal information. Whereas levitation and deposition occur for strongly repulsive or attractive potentials, association is exponentially sensitive to weak interactions influenced by adsorbed layer architectures and surface heterogeneity. Systematic experiments reveal how BSA orientation and PEO molecular weight produce adsorbed layers that either conceal or expose substrate heterogeneities to generate a continuum of colloid-surface association lifetimes. These measurements provide simultaneous access to a broad range of information that consistently indicates purely repulsive BSA and PEO interactions and a role for surface heterogeneity in colloid-surface association. The demonstrated capability to measure nonspecific protein interactions provides a basis for future measurements of specific protein interactions.
Everett
W Neil
WN
Department of Mechanical Engineering, Texas A&M University, College Station, Texas 77843, USA.
Wu
Hung-Jen
HJ
Anekal
Samartha G
SG
Sue
Hung-Jue
HJ
Bevan
Michael A
MA
D003102:Colloids / Q000737:chemistry*; D004058:Diffusion; D007700:Kinetics; D046911:Macromolecular Substances / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D015336:Molecular Probe Techniques; D011092:Polyethylene Glycols / Q000737:chemistry*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000648:ultrastructure*
D003102:Colloids; D046911:Macromolecular Substances; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols