Orientation of Biotin-Binding Sites in Streptavidin Adsorbed onto the Surface of Polythiophene Films.
Kamil AwsiukPanagiota PetrouAngelos ThanassoulasJoanna RaczkowskaPublished in: Langmuir : the ACS journal of surfaces and colloids (2019)
The orientation of biotin-binding sites of streptavidin adsorbed to thin films of three polythiophenes (PTs), namely, regioregular poly(3-hexylthiophene) (RP3HT), regiorandom poly(3-butylthiophene) (P3BT), and poly(3,3‴-didodecylquaterthiophene) (PQT12), has been investigated. Polymer films were examined prior to and after protein adsorption with atomic force microscopy and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Principal component analysis (PCA) applied to ToF-SIMS data revealed subtle changes in surface chemistry of polymer films and orientation of adsorbed streptavidin. PCA resolved the surface alignment of alkyl side chains and differentiated the ToF-SIMS data for PQT12, RP3HT, and P3BT, verifying an amorphous morphology for P3BT and a semicrystalline one for PQT12 and RP3HT. After the characterization of the polymeric films, streptavidin adsorption from solutions with different protein concentrations (up to 300 μg/mL) has been conducted. The PCA results distinguished between amino acids characteristic for external regions of streptavidin molecules adsorbed to different PTs suggest that streptavidin adsorbed to PQT12 exposes molecular regions rich in tryptophan and tyrosine, which are components of the biotin-binding sites. The latter results were confirmed using biotin-labeled horse radish peroxidase to estimate the exposed binding sites of streptavidin adsorbed onto the different PT films. The analysis of streptavidin structure suggests that interaction between polythiophene film and dipole moment of streptavidin subunit is responsible for orientation of biotin-binding sites.
Keyphrases
- mass spectrometry
- room temperature
- atomic force microscopy
- ms ms
- amino acid
- drug delivery
- carbon nanotubes
- single molecule
- high speed
- ionic liquid
- nitric oxide
- machine learning
- cancer therapy
- hydrogen peroxide
- gold nanoparticles
- reduced graphene oxide
- computed tomography
- pet imaging
- simultaneous determination
- drug release