AVS2013 Session BA+AI+AS+BI+IS+NL-MoM: Biomolecules at Aqueous Interfaces

Monday, October 28, 2013 8:20 AM in Room 203 A

Monday Morning

Time Period MoM Sessions | Abstract Timeline | Topic BA Sessions | Time Periods | Topics | AVS2013 Schedule

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8:20 AM BA+AI+AS+BI+IS+NL-MoM-1 Selected Studies of Biomolecular Interactions
Kenneth B. Eisenthal, Benjamin Doughty, Yi Rao, Samuel Kazer, Sheldon Kwok, Nicholas Turro (Columbia University)

The work reported here utilizes a sensitive method for the investigation of biomolecular interactions that has the important characteristic of not requiring chemical labels, e.g. fluorophores, nor invasive detection methods. The surface selective second order spectroscopies, second harmonic, SHG, and sum frequency generation, SFG, allow one to probe equilibrium properties and time dependent changes in the electronic and vibrational structure of molecules located at interfaces. In addition they have the special feature of being able to monitor changes in the electrical charge of the interacting molecules. 1) A new way is presented to measure the binding constants of molecules, e.g. drugs and proteins, with DNA tethered to colloidal microparticles suspended in aqueous solution. 2) Time resolved second harmonic generation was used to observe the binding of an enzyme to its recognition site on DNA, followed by the cleaving of DNA into a small and a large fragment, and the subsequent DNA rehybridization dynamics. 3) The relative orientation of two molecules bound to DNA is manipulated by changing the number of nucleotide base pairs separating them. The interference between the SH electric fields generated by the pair of molecules is modulated because their relative orientation changes as the number of nucleotide base pairs separating them is changed. With this method we have a new way to probe structural changes in DNA due to the binding of biomolecules to it.

9:00 AM BA+AI+AS+BI+IS+NL-MoM-3 Probing Nanoparticle-lipid Bilayer Interactions with Nonlinear Optics
Franz Geiger (Northwestern University)
The interaction of engineered nanoparticles with biological membranes is an important and necessary first step for cellular uptake. Here, we probe this interaction by applying second harmonic and vibrational sum frequency generation as well as the Eisenthal chi(3) method to supported bilayer-based model systems as well as shewanella and daphnia magna, chosen as important biological endpoints, exposed to 4 nm sized noble metal nanoparticles surrounded by negatively and positively charged ligands. Our studies are complemented by a plethora of supporting experiments based on quartz crystal microbalance, zeta potential, and related experiments. We find that Coulomb's law dictates much of the interactions in the particular systems studied here.
9:40 AM BA+AI+AS+BI+IS+NL-MoM-5 Characterizing the Protein-Surface Interactions that Control Diatom Biomineralization
Joe Baio (Oregon State University); Mischa Bonn, Tobias Weidner (Max Planck Institute for Polymer Research, Germany)

The assembly of mineralized tissues can be initiated and controlled by proteins. One such system, is the formation of silica-based cell walls in marine, single celled organisms, where biomineralization is regulated by protein−mineral interactions. The diatom species Cylindrotheca fusiformis assembles supramolecular silica structures via proteins called sillafins. In a silicic acid solution, specific repeat units within this protein, SSKKSGSYSGSKGSKRRIL (R5), induce the formation of silica-protein composite nanoparticles. The protein−surface interaction that drives self-assembly is likely controlled by both the secondary structural motifs of the protein and specific contacts between the surface atoms and key protein side chains. In this study, we characterized the R5-SiO2 interactions that drive this self-assembly process by both near edge x-ray absorption fine structure (NEXAFS) spectroscopy and in situ sum frequency generation (SFG) spectroscopy. Two peaks within the amide I vibrational band of the SFG spectra, 1640 and 1670 cm-1, indicate that the R5 peptide retains a beta sheet conformation when interacting with SiO2. Expanding upon this characterization of secondary structure, the introduction of isotopic labeled amino acids within the peptide allowed us to probe the orientations of individual side chains by SFG. This SFG characterization was complemented by the observed polarization dependence of the NEXAFS C1s to π* transition which provided details of the binding geometry of the single tyrosine within R5.

10:00 AM BA+AI+AS+BI+IS+NL-MoM-6 Probing the Effects of Different Ions on the Formation of Microstructure Within Collagen Hydrogels by Second Harmonic Generation (SHG) Microscopy
Xuye Lang, Julia G. Lyubovitsky (University of California, Riverside)

In this study we aimed to explore the nucleation, assembly and the 3-D microstructure of collagen hydrogels in situ with second harmonic generation (SHG) microscopy. Transmission electron microscopy (TEM) and optical density (OD) were carried out as well in order to complement the SHG measurements. The goal was to generate the knowledge to accelerate rational design of collagen-based biomedical products. In this work, we employed 0, 150, 300, 600, 900 mM NaCl concentrations and in a separate experiment 0, 5, 10, 20, 50, 75, 100, 150, 300 mM Na2SO4 concentrations of salts needed for the assembly of collagen hydrogels. Specifically, we characterized collagen hydrogels prepared from 2 g/l and 4 g/l initial collagen concentrations as well as several incubation temperatures. For samples incubated with NaCl, incubation under the room temperature (RT) and 27°C gave similar OD values. These OD values were higher than the OD values for 37°C incubated samples. Delay time became shorter upon elevating the polymerization temperature. For samples incubated with Na2SO4, there were two regimes for collagen polymerization, Na2SO4 concentration 5 mM – 50 mM and 100 mM -300 mM. Fibers were longer when NaCl concentration was 150-600 mM compared to 0 and 900 mM NaCl for both collagen concentrations under RT, 27°C and 37°C. In general, fibers were small when incubated at 37°C compared to fibers formed under RT and 27°C. TEM measurement showed that there were collagen fibers with a characteristic striation structure in all collagen and NaCl concentrations. The fibrils exhibited a twisted morphology in 2 g/l collagen hydrogels.

10:20 AM BREAK
10:40 AM BA+AI+AS+BI+IS+NL-MoM-8 Sum Frequency Generation (SFG) Vibrational Spectroscopy Studies of Molecules at Solid-Liquid and Solid-Gas Interfaces
Gabor Somorjai (University of California, Berkeley and Lawrence Berkeley National Laboratory); Xiaojun Cai (Lawrence Berkeley National Laboratory)

Construction of a femtosecond broad-band laser enables us to simultaneously monitor the CH and CO vibrational SFG spectra, thus allowing more actual characterization of reaction intermediates at solid-liquid and solid-gas interfaces. Using a picosecond laser we compare the spectroscopy using these two different laser systems and they will be discussed in some detail. In addition, sum frequency generation vibrational spectroscopy, high-pressure scanning tunneling microscopy and ambient-pressure X-ray photoelectron spectroscopy as well as other synchrotron-based techniques (X-ray adsorption) that enable the investigation of surfaces under reaction conditions on the atomic and molecular level will be reviewed.

We investigate solid-liquid and solid-solid interfaces (buried interfaces) as they adsorb and react with diatomic and organic molecules in dynamic state at various pressures and temperatures.

11:20 AM BA+AI+AS+BI+IS+NL-MoM-10 A Molecular View of Water Interacting with Climate-active Ice Nucleating Proteins
Ravindra Pandey (Max Planck Institute for Polymer Research, Germany); Janine Fröhlich, Ulrich Pöschl (Max Planck Institute for Chemistry, Germany); Mischa Bonn, Tobias Weidner (Max Planck Institute for Polymer Research, Germany)
Specific bacteria, such as Pseudomonas syringae, effectively attack plants by using ice-nucleating proteins anchored to their outer cell surfaces. Ice nucleating proteins promote the local crystallization of ice at temperatures that would otherwise not allow ice formation. The frost damage caused by ice crystals then facilitates bacterial invasion of the affected plants. Ice nucleating proteins not only play an important role for agriculture, but are also very important for atmospheric processes: airborne ice-nucleating proteins have been shown to be among the most effective promoters of ice particle formation in the atmosphere. A recent survey of microorganisms in the troposphere biome by NASA has discovered massive emissions of biogenic ice nucleators from large forest areas like the amazon, which likely change precipitation patterns and may affect the global climate. To understand biogenic ice formation, a detailed molecular level picture of the mechanism by which ice-nucleating proteins interact with water molecules is important. Sum frequency generation (SFG) spectroscopy – owing to its inherent interface sensitivity – is ideally suited to determine the structure and dynamics of water molecules at interfaces. We have investigated the interaction a monolayer of the ice-nucleating protein inaZ with water using static and time-resolved SFG spectroscopy. When cooling the sample from room temperature to near-freezing temperatures (~5°C for D2O), inaZ significantly increases the structural order of water molecules in contact with inaZ proteins. This effect was not observed for liquid water surfaces without the protein or for protein monolayers which are not ice nucleators. SFG spectra in the CH and the amide I region also indicated a change of protein structure near the nucleation temperature. Femtosecond, time-resolved 2-dimensional SFG spectroscopy is used to quantify the heterogeneity of protein-bound water molecules and their structural dynamics.
Time Period MoM Sessions | Abstract Timeline | Topic BA Sessions | Time Periods | Topics | AVS2013 Schedule