Capturing protein dynamics with time-resolved luminescence spectroscopy

The presented doctoral research utilizes time-resolved spectroscopy to characterize protein dynamics and folding mechanisms. We resolve millisecond-timescale folding by coupling time-resolved fluorescence energy transfer (trFRET) to a continuous flow microfluidic mixer to obtain intramolecular distance distributions throughout the folding process. We have elucidated the folding mechanisms of two cytochromes---one that exhibits two-state folding (cytochrome cb₅₆₂) and one that has both a kinetic refolding intermediate ensemble and a distinct equilibrium unfolding intermediate (cytochrome c₅₅₂). Our data reveal that the distinct structural features of cytochrome c₅₅₂ contribute to its thermostability.

We have also investigated intrachain contact dynamics in unfolded cytochrome cb₅₆₂ by monitoring electron transfer, which occurs as the heme collides with a ruthenium photosensitizer, covalently bound to residues along the polypeptide. Intrachain diffusion for chemically denatured proteins proceeds on the microsecond timescale with an upper limit of 0.1 microseconds. The power-law dependence (slope = -1.5) of the rate constants on the number of peptide bonds between the heme and Ru complex indicate that cytochrome cb₅₆₂ is minimally frustrated.

In addition, we have explored the pathway dependence of electron tunneling rates between metal sites in proteins. Our research group has converted cytochrome b₅₆₂ to a c-type cytochrome with the porphyrin covalently bound to cysteine sidechains. We have investigated the effects of the changes to the protein structure (i.e., increased rigidity and potential new equatorial tunneling pathways) on the electron transfer rates, measured by transient absorption, in a series of ruthenium photosensitizer-modified proteins.

1. Astronomical photometry at wavelengths of 8.5, 10.5, and 11.6 µm. 2. Infrared emission from asteroids at wavelengths of 8.5, 10.5, and 11.6 µm

The purpose of this thesis is to present new observations of thermal-infrared radiation from asteroids. Stellar photometry was performed to provide standards for comparison with the asteroid data. The details of the photometry and the data reduction are discussed in Part 1. A system of standard stars is derived for wavelengths of 8.5, 10.5 and 11.6 µm and a new calibration is adopted. Sources of error are evaluated and comparisons are made with the data of other observers.

The observations and analysis of the thermal-emission observations of asteroids are presented in Part 2. Thermal-emission lightcurve and phase effect data are considered. Special color diagrams are introduced to display the observational data. These diagrams are free of any model-dependent assumptions and show that asteroids differ in their surface properties.

On the basis of photometric models, (4) Vesta is thought to have a bolometric Bond albedo of about 0.1, an emissivity greater than 0.7 and a true radius that is close to the model value of 300^(+50)_(-30)km. Model albedos and model radii are given for asteroids 1, 2, 4, 5, 6, 7, 15, 19, 20, 27, 39, 44, 68, 80, 324 and 674. The asteroid (324) Bamberga is extremely dark with a model (~bolometric Bond) albedo in the 0.01 - 0.02 range, which is thought to be the lowest albedo yet measured for any solar-system body. The crucial question about such low-albedo asteroids is their number and the distribution of their orbits.

Schottky barriers on compound semiconductors: I. HgSe highly electronegative contacts. II. Au Schottky barriers on n-Ga_(1-x)Al_xAs

I. HgSe is deposited on various semiconductors, forming a semimetal/semiconductor "Schottky barrier" structure. Polycrystalline, evaporated HgSe produces larger Schottky barrier heights on n-type semiconductors than does Au, the most electronegative of the elemental metals. The barrier heights are about 0.5 eV greater than those of Au on ionic semiconductors such as ZnS, and 0.1 to 0.2 eV greater for more covalently bonded semiconductors. A novel structure,which is both a lattice matched heterostructure and a Schottky barrier, is fabricated by epitaxial growth of HgSe on CdSe using hydrogen transport CVD. The Schottky barrier height for this structure is 0.73 ± 0.02 eV, as measured by the photoresponse method. This uncertainty is unusually small; and the magnitude is greater by about a quarter volt than is achievable with Au, in qualitative agreement with ionization potential arguments.

II . The Schottky barrier height of Au on chemically etched n-Ga_1-x Al_xAs was measured as a function of x. As x increases, the barrier height rises to a value of about 1.2 eV at x ≈ 0.45 , then decreases to about 1.0 eV as x approaches 0.83. The barrier height deviates in a linear way from the value predicted by the "common anion" rule as the AlAs mole fraction increases. This behavior is related to chemical reactivity of the Ga_1-x Al_xAs surface.

PP backward elastic scattering between 0.70 and 2.37 GeV/c

̄pp backward elastic scattering has been measured for the cos θ_cm region between – 1.00 and – 0.88 and for the incident ̄p laboratory momentum region between 0.70 and 2.37 GeV/c. These measurements, done in intervals of approximately 0.1 GeV/c, have been performed at the Alternating Gradient Synchrotron at Brookhaven National Laboratory during the winter of 1968. The measured differential cross sections, binned in cos θ_cm intervals of 0.02, have statistical errors of about 10%. Backward dipping exists below 0.95 GeV/c and backward peaking above 0.95 GeV/c. The 180˚ differential cross section extrapolated from our data shows a sharp dip centered at 0.95 GeV/c and a broad hump centered near 1.4 GeV/c. Our data have been interpreted in terms of resonance effects and in terms of diffraction dominance effects.