Polyvalent display and packing of peptides and proteins on semiconductor quantum dots: predicted versus experimental results.
Quantum dots (QDs) are loaded with a series of peptides and proteins of increasing size, including a <20 residue peptide, myoglobin, mCherry, and maltose binding protein, which together cover a range of masses from <2.2 to approximately 44 kDa. Conjugation to the surface of dihydrolipoic acid-functionalized QDs is facilitated by polyhistidine metal affinity coordination. Increasing ratios of dye-labeled peptides and proteins are self-assembled to the QDs and then the bioconjugates are separated and analyzed using agarose gel electrophoresis. Fluorescent visualization of both conjugated and unbound species allows determination of an experimentally derived maximum loading number. Molecular modeling utilizing crystallographic coordinates or space-filling structures of the peptides and proteins also allow the predicted maximum loadings to the QDs to be estimated. Comparison of the two sets of results provides insight into the nature of the QD surface and reflects the important role played by the nanoparticle's hydrophilic solubilizing surface ligands. It is found that for the larger protein molecules steric hindrance is the major packing constraint. In contrast, for the smaller peptides, the number of available QD binding sites is the principal determinant. These results can contribute towards an overall understanding of how to engineer designer bioconjugates for both QDs and other nanoparticle materials.
Prasuhn
Duane E
DE
Center for Bio/Molecular Science and Engineering, Code 6900, U.S. Naval Research Laboratory, Washington, DC 20375, USA.
Deschamps
Jeffrey R
JR
Susumu
Kimihiro
K
Stewart
Michael H
MH
Boeneman
Kelly
K
Blanco-Canosa
Juan B
JB
Dawson
Philip E
PE
Medintz
Igor L
IL
D004587:Electrophoresis, Agar Gel; D008958:Models, Molecular; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D045663:Quantum Dots; D012666:Semiconductors; D008063:Thioctic Acid / Q000031:analogs & derivatives / Q000737:chemistry
D010455:Peptides; D011506:Proteins; D008063:Thioctic Acid; C015144:dihydrolipoic acid