How Ubiquitin Unfolds after Transfer into the Gas Phase

The structural evolution of ubiquitin after transfer into the gas phase was studied by electron capture dissociation. Site-specific fragment yields show that ubiquitin’s solution fold is overall unstable in the gas phase, but unfolding caused by loss of solvent is slowest in regions stabilized by salt bridges

Activated Ion Electron Capture Dissociation (AI ECD) of proteins: synchronization of infrared and electron irradiation with ion magnetron motion.

Here, we show that to perform activated ion electron capture dissociation (AI-ECD) in a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer equipped with a CO(2) laser, it is necessary to synchronize both infrared irradiation and electron capture dissociation with ion magnetron motion. This requirement is essential for instruments in which the infrared laser is angled off-axis, such as the Thermo Finnigan LTQ FT. Generally, the electron irradiation time required for proteins is much shorter (ms) than that required for peptides (tens of ms), and the modulation of ECD, AI ECD, and infrared multiphoton dissociation (IRMPD) with ion magnetron motion is more pronounced. We have optimized AI ECD for ubiquitin, cytochrome c, and myoglobin; however the results can be extended to other proteins. We demonstrate that pre-ECD and post-ECD activation are physically different and display different kinetics. We also demonstrate how, by use of appropriate AI ECD time sequences and normalization, the kinetics of protein gas-phase refolding can be deconvoluted from the diffusion of the ion cloud and measured on the time scale longer than the period of ion magnetron motion

Topoisomer Differentiation of Molecular Knots by FTICR MS: Lessons from Class II Lasso Peptides

Lasso peptides constitute a class of bioactive peptides sharing a knotted structure where the C-terminal tail of the peptide is threaded through and trapped within an N-terminalmacrolactamring. The structural characterization of lasso structures and differentiation from their unthreaded topoisomers is not trivial and generally requires the use of complementary biochemical and spectroscopic methods. Here we investigated two antimicrobial peptides belonging to the class II lasso peptide family and their corresponding unthreaded topoisomers: microcin J25 (MccJ25), which is known to yield two-peptide product ions specific of the lasso structure under collisioninduced dissociation (CID), and capistruin, for which CID does not permit to unambiguously assign the lasso structure. The two pairs of topoisomers were analyzed by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) upon CID, infrared multiple photon dissociation (IRMPD), and electron capture dissociation (ECD). CID and ECDspectra clearly permitted to differentiate MccJ25 from its non-lasso topoisomer MccJ25-Icm, while for capistruin, only ECD was informative and showed different extent of hydrogen migration (formation of c\bullet/z from c/z\bullet) for the threaded and unthreaded topoisomers. The ECD spectra of the triply-charged MccJ25 and MccJ25-lcm showed a series of radical b-type product ions {\eth}b0In{\TH}. We proposed that these ions are specific of cyclic-branched peptides and result from a dual c/z\bullet and y/b dissociation, in the ring and in the tail, respectively. This work shows the potentiality of ECD for structural characterization of peptide topoisomers, as well as the effect of conformation on hydrogen migration subsequent to electron capture

The Role of the D13 (1520) Resonance in eta Electroproduction

We investigate the electroproduction of eta mesons below a center of momentum energy of 1.6 GeV, with particular emphasis on the roles of the N*(1535) and N*(1520) resonances. Using the effective Lagrangian approach, we show that the transverse helicity amplitude of the N*(1535) can be extracted with good accuracy from the new eta electroproduction data, under reasonable assumptions for the strength of the longitudinal helicity amplitude. In addition, although the differential cross section is found to to have a small sensitivity to the N*(1520) resonance, it is shown that a recently completed double polarization experiment is very sensitive to this resonance.Comment: 7 pages, Revtex, 3 figure

A hydrogen beam to characterize the ASACUSA antihydrogen hyperfine spectrometer

The antihydrogen programme of the ASACUSA collaboration at the antiproton decelerator of CERN focuses on Rabi-type measurements of the ground-state hyperfine splitting of antihydrogen for a test of the combined Charge-Parity-Time symmetry. The spectroscopy apparatus consists of a microwave cavity to drive hyperfine transitions and a superconducting sextupole magnet for quantum state analysis via Stern-Gerlach separation. However, the small production rates of antihydrogen forestall comprehensive performance studies on the spectroscopy apparatus. For this purpose a hydrogen source and detector have been developed which in conjunction with ASACUSA's hyperfine spectroscopy equipment form a complete Rabi experiment. We report on the formation of a cooled, polarized, and time modulated beam of atomic hydrogen and its detection using a quadrupole mass spectrometer and a lock-in amplification scheme. In addition key features of ASACUSA's hyperfine spectroscopy apparatus are discussed.

Electron Capture Dissociation Mass Spectrometry of Tyrosine Nitrated Peptides

In vivo protein nitration is associated with many disease conditions that involve oxidative stress and inflammatory response. The modification involves addition of a nitro group at the position ortho to the phenol group of tyrosine to give 3-nitrotyrosine. To understand the mechanisms and consequences of protein nitration, it is necessary to develop methods for identification of nitrotyrosine-containing proteins and localization of the sites of modification.Here, we have investigated the electron capture dissociation (ECD) and collision-induced association (CID) behavior of 3-nitrotyrosine-containing peptides. The presence of nitration did not affect the CID behavior of the peptides. For the doubly-charged peptides, addition of nitration severely inhibited the production of ECD sequence fragments. However, ECD of the triply-charged nitrated peptides resulted in some singly-charged sequence fragments. ECD of the nitrated peptides is characterized by multiple losses of small neutral species including hydroxyl radicals, water and ammonia. The origin of the neutral losses has been investigated by use of activated ion (AI) ECD. Loss of ammonia appears to be the result of non-covalent interactions between the nitro group and protonated lysine side-chains

Threshold eta and eta' electroproduction off nucleons

The electroproduction of eta and eta' mesons on the proton and the neutron is investigated at tree level within the framework of U(3) chiral perturbation theory. In addition to the Born terms low-lying resonances such as the vector mesons and J^P= 1/2^+, 1/2^- baryon resonances are included explicitly and their contributions are calculated. Results for the separated differential cross sections are presented.Comment: 24 pages, 7 figure

Quasifree Pion Electroproduction from Nuclei in the Δ\Delta Region

We present calculations of the reaction $A(e,e^\prime \pi N)B$ in the distorted wave impulse approximation. The reaction allows for the study of the production process in the nuclear medium without being obscured by the details of nuclear transition densities. First, a pion electroproduction operator suitable for nuclear calculations is obtained by extending the Blomqvist-Laget photoproduction operator to the virtual photon case. The operator is gauge invariant, unitary, reference frame independent, and describes the existing data reasonably well. Then it is applied in nuclei to predict nuclear cross sections under a variety of kinematic arrangements. Issues such as the effects of gauge-fixing, the interference of the $\Delta$ resonance with the background, sensitivities to the quadrupole component of the $\Delta$ excitation and to the electromagnetic form factors, the role of final-state interactions, are studied in detail. Methods on how to experimentally separate the various pieces in the coincidence cross section are suggested. Finally, the model is compared to a recent SLAC experiment.Comment: 27 pages in REVTEX, plus 22 PS figures embedded using psfig.sty (included), uuencode

The relativistic impulse approximation for the exclusive electrodisintegration of the deuteron

The electrodisintegration of the deuteron in the frame of the Bethe-Salpeter approach with a separable kernel of the nucleon-nucleon interaction is considered. This conception keeps the covariance of a description of the process. A comparison of relativistic and nonrelativistic calculations is presented. The factorization of the cross section of the reaction in the impulse approximation is obtained by analytical calculations. It is shown that the photon-neutron interaction plays an important role.Comment: 31 pages, 14 figures, 1 tabl