Shake-up effect in photoluminescence of integer quantum Hall system formed in InGaAs/InP quantum wells.

Polarization-resolved magneto-photoluminescence is studied in InGaAs/InP single quantum wells. In the range of the filling factor ν ≥ 4 the number of populated Landau levels contributing to the photoluminescence is found to be equal to the corresponding filling factor, while at ν ≤ 3 the number of emitting Landau levels is larger than the filling factor, which implies an occupancy of the Landau levels above the Fermi level. Such partial occupancy of the Landau levels with energies higher than the Fermi energy is due to shake-up processes caused by electron-electron interaction. In accordance to the theory, at the filling factor around ν = 2 the shake-up process was found to manifest itself in the downshifted cusp of the energy of the σ- polarized emission from the excited Landau levels, while no change was observed in the energy of the σ+ polarized emission. The different energies of differently spin-polarized excited Landau levels cause the magnetic field induced polarization of the emission from excited Landau levels. In addition, the bound electron-hole excitonic complexes (trions) associated with different Landau levels were observed.

Magnetic field driven interminiband charge transfer in InGaAs/InP superlattices.

The characteristic energies, occupancies and polarizations of the minibands formed by the Γ-Γ and Γ-Xz interlayer electon tunnelings in the InGaAs/InP superlattices are studied in the regime of the integer quantum Hall effect by polarization resolved photoluminescence. Accordingly, the magnetic field induced shrinkage of the interminiband gap, predicted by the theory, and as a consequence, the redistribution of charge over the superlattice minibands and the depolarization of the quantum Hall electron states are observed at odd filling factors. The response of the electrons residing in the InGaAs/InP superlattice minibands to the magnetic field is found very similar to the corresponding response of the electrons confined in the symmetric and anti-symmetric two-dimensional minibands of GaAs/AlGaAs double quantum wells. The presented results are evidence of the formation of the correlated states in multi-component electron systems formed in semiconductor multiple layers at odd filling factors.

Probing molecular interactions in intact antibody: antigen complexes, an electrospray time-of-flight mass spectrometry approach.

Using a combination of nanoflow-electrospray ionization and time-of-flight mass spectrometry we have analyzed the oligomeric state of the recombinant V antigen from Yersinia pestis, the causative agent of plague. The mass spectrometry results show that at pH 6.8 the V antigen in solution exists predominantly as a dimer and a weakly associated tetramer. A monoclonal antibody 7.3, raised against the V antigen, gave rise to mass spectra containing a series of well-resolved charge states at m/z 6000. After addition of aliquots of solution containing V antigen in substoichiometric and molar equivalents, the spectra revealed that two molecules of the V antigen bind to the antibody. Collision-induced dissociation of the antibody-antigen complex results in the selective release of the dimer from the complex supporting the proposed 1:2 antibody:antigen stoichiometry. Control experiments with the recombinant F1 antigen, also from Yersinia pestis, establish that the antibody is specific for the V antigen because no complex with F1 was detected even in the presence of a 10-fold molar excess of F1 antigen. More generally this work demonstrates a rapid means of assessing antigen subunit interactions as well as the stoichiometry and specificity of binding in antibody-antigen complexes.

Macromolecular organization of the Yersinia pestis capsular F1 antigen: insights from time-of-flight mass spectrometry.

Mass spectrometry has been used to examine the subunit interactions in the capsular F1 antigen from Yersinia pestis, the causative agent of the plague. Introducing the sample using nanoflow electrospray from solution conditions in which the protein remains in its native state and applying collisional cooling to minimize the internal energy of the ions, multiple subunit interactions have been maintained. This methodology revealed assemblies of the F1 antigen that correspond in mass to both 7-mers and 14-mers, consistent with interaction of two seven-membered units. The difference between the calculated masses and those measured experimentally for these higher-order oligomers was found to increase proportionately with the size of the complex. This is consistent with a solvent-filled central cavity maintained on association of the 7-mer to the 14-mer. The charge states of the ions show that an average of one and four surface accessible basic side-chains are involved in maintaining the interactions between the 7-mer units and neighboring subunits, respectively. Taken together, these findings provide new information about the stoichiometry and packing of the subunits involved in the assembly of the capsular antigen structure. More generally, the data show that the symmetry and packing of macromolecular complexes can be determined solely from mass spectrometry, without any prior knowledge of higher order structure

Observation of the noncovalent assembly and disassembly pathways of the chaperone complex MtGimC by mass spectrometry.

We have analyzed a newly described archaeal GimC/prefoldin homologue, termed MtGimC, by using nanoflow electrospray coupled with time-of-flight MS. The molecular weight of the complex from Methanobacterium thermoautotrophicum corresponds to a well-defined hexamer of two alpha subunits and four beta subunits. Dissociation of the complex within the gas phase reveals a quaternary arrangement of two central subunits, both alpha, and four peripheral beta subunits. By constructing a thermally controlled nanoflow device, we have monitored the thermal stability of the complex by MS. The results of these experiments demonstrate that a significant proportion of the MtGimC hexamer remains intact under low-salt conditions at elevated temperatures. This finding is supported by data from CD spectroscopy, which show that at physiological salt concentrations, the complex remains stable at temperatures above 65 degrees C. Mass spectrometric methods were developed to monitor in real time the assembly of the MtGimC hexamer from its component subunits. By using this methodology, the mass spectra recorded throughout the time course of the experiment showed the absence of any significantly populated intermediates, demonstrating that the assembly process is highly cooperative. Taken together, these data show that the complex is stable under the elevated temperatures that are appropriate for its hyperthermophile host and demonstrate that the assembly pathway leads exclusively to the hexamer, which is likely to be a structural unit in vivo.