Multiple Evolutionary Origins of Ubiquitous Cu2+ and Zn2+ Binding in the S100 Protein Family

The S100 proteins are a large family of signaling proteins that play critical roles in biology and disease. Many S100 proteins bind Zn2+, Cu2+, and/or Mn2+ as part of their biological functions; however, the evolutionary origins of binding remain obscure. One key question is whether divalent transition metal binding is ancestral, or instead arose independently on multiple lineages. To tackle this question, we combined phylogenetics with biophysical characterization of modern S100 proteins. We demonstrate an earlier origin for established S100 subfamilies than previously believed, and reveal that transition metal binding is widely distributed across the tree. Using isothermal titration calorimetry, we found that Cu2+ and Zn2+ binding are common features of the family: the full breadth of human S100 paralogs—as well as two early-branching S100 proteins found in the tunicate Oikopleura dioica—bind these metals with μM affinity and stoichiometries ranging from 1:1 to 3:1 (metal:protein). While binding is consistent across the tree, structural responses to binding are quite variable. Further, mutational analysis and structural modeling revealed that transition metal binding occurs at different sites in different S100 proteins. This is consistent with multiple origins of transition metal binding over the evolution of this protein family. Our work reveals an evolutionary pattern in which the overall phenotype of binding is a constant feature of S100 proteins, even while the site and mechanism of binding is evolutionarily labile

Eye lens β-crystallins are predicted by native ion mobility-mass spectrometry and computations to form compact higher-ordered heterooligomers

Eye lens crystallin proteins maintain the refractive properties of the lens but are not replaced after denucleation. Rolland et al. use native ion mobility-mass spectrometry, kinetics experiments, and computations to reveal that b-crystallins form heterodimers. These likely assemble into compact heterooligomers that enable the very high protein concentrations found in lens tissue

Direct and Simultaneous Observation of Ultrafast Electron and Hole Dynamics in Germanium

Understanding excited carrier dynamics in semiconductors is crucial for the development of photovoltaics and efficient photonic devices. However, overlapping spectral features in optical/NIR pump-probe spectroscopy often render assignments of separate electron and hole carrier dynamics ambiguous. Here, ultrafast electron and hole dynamics in germanium nanocrystalline thin films are directly and simultaneously observed by attosecond transient absorption spectroscopy (ATAS) in the extreme ultraviolet at the germanium M_{4,5}-edge (~30 eV). We decompose the ATAS spectra into contributions of electronic state blocking and photo-induced band shifts at a carrier density of 8*10^{20}cm^{-3}. Separate electron and hole relaxation times are observed as a function of hot carrier energies. A first order electron and hole decay of ~1 ps suggests a Shockley-Read-Hall recombination mechanism. The simultaneous observation of electrons and holes with ATAS paves the way for investigating few to sub-femtosecond dynamics of both holes and electrons in complex semiconductor materials and across junctions.Comment: Includes Supplementary Informatio

Averages of b-hadron Properties at the End of 2005

This article reports world averages for measurements on b-hadron properties obtained by the Heavy Flavor Averaging Group (HFAG) using the available results as of at the end of 2005. In the averaging, the input parameters used in the various analyses are adjusted (rescaled) to common values, and all known correlations are taken into account. The averages include lifetimes, neutral meson mixing parameters, parameters of semileptonic decays, branching fractions of B meson decays to final states with open charm, charmonium and no charm, and measurements related to CP asymmetries

Observation of the Dynamic Beta Effect at CESR with CLEO

Using the silicon strip detector of the CLEO experiment operating at the Cornell Electron-positron Storage Ring (CESR), we have observed that the horizontal size of the luminous region decreases in the presence of the beam-beam interaction from what is expected without the beam-beam interaction. The dependence on the bunch current agrees with the prediction of the dynamic beta effect. This is the first direct observation of the effect.Comment: 9 page uuencoded postscript file, postscritp file also available through http://w4.lns.cornell.edu/public/CLNS, submitted to Phys. Rev.

Protonation Isomers of Highly Charged Protein Ions Can Be Separated in FAIMS-MS

High-field asymmetric waveform ion mobility spectrometry-mass spectrometry (FAIMS-MS) can resolve over an order of magnitude more conformers for a given protein ion than alternative methods. Such an expansion in separation space results, in part, from protein ions with masses of \u3e29 kDa undergoing dipole alignment in the high electric field of FAIMS, and the resolution of ions that adopt pendular vs free rotor states. In this study, FAIMS-MS, collision-induced dissociation (CID), and travelling wave (TW) IMS-MS were used to investigate the pendular and free rotor states of protonated carbonic anhydrase II (CAII, 29 kDa). The electrospray ionization additive 1,2-butylene carbonate was used to increase protein charge states and ensure extended ion conformations were formed. For relatively high charge states in which dipole alignment occurs (30e38þ), FAIMS-MS can baseline resolve the isobaric pendular and free rotor ion populations. For TWIMS-MS, these same charge states resulted in monomodal arrival time distributions with collision cross sections corresponding to highly extended ion conformations. Interestingly, CID of FAIMS-selected pendular and free rotor ion populations resulted in significantly different frag-mentation patterns. For example, CID of the dipole aligned CAII 37þ resulted in cleavages C-terminal to residue 183, 192 and 196, whereas cleavage sites for the free rotor population occurred near residues 12 and 238. Given that the cleavage sites are ’directed’ by protonation sites in the CID of protein ions, and highly charged protein ions adopt extended conformations with the same or very similar collision cross sections, these results indicate that the pendular and free rotor populations separated in FAIMS can be attributed to protonation isomers. Moreover, the extent of protein ion charging in FAIMS-MS decreased substantially as the carrier gas flow rate decreased, indicating that ion charging in FAIMS-MS can be limited by proton-transfer reactions. Given that the total mass of proton charge carriers corresponds to less than 0.2% the mass of CAII, we anticipate that FAIMS-MS can be used to separate intact isobaric proteoforms with masses of at least ~29 kDa that result from alternative sites of post-translational modifications

Further Search for the Two-Photon Production of the Glueball Candidate fJ(2220)f_{J}(2220)

The CLEOII detector at the Cornell e+ e- storage ring CESR has been used to search for the two-photon production of the $f_J(2220)$ decaying into pi+ pi-. No evidence for a signal is found in data corresponding to an integrated luminosity of 4.77/fb and a 95% CL upper limit on $\Gamma_{two-photon} * BR{pi+ pi-}$ of 2.5 eV is set. If this result is combined with the BES Collaboration's measurement of $f_J(2220) -> pi+ pi-$ in radiative $J/\psi$ decay, a 95% CL lower limit on the stickiness of the $f_J(2220)$ of 73 is obtained. If the recent CLEO result for \Gamma_{two-photon} * BR{\K_S K_S} is combined with the present result, the stickiness of the $f_J(2220)$ is found to be larger than 102 at the 95% CL. These results for the stickiness (the ratio of the probabilities for two-gluon coupling and two-photon coupling) provide further support for a substantial neutral parton content in the $f_J(2220)$.Comment: 8 pages, postscript file also available through http://w4.lns.cornell.edu/public/CLN

Resonant Structure of τ→3ππ0ντ\tau\to 3\pi\pi^{0}\nu_{\tau} and τ→ωπντ\tau\to \omega\pi\nu_{\tau} Decays

The resonant structure of the four pion final state in the decay $\tau \to 3\pi\pi^0\nu_\tau$ is analyzed using 4.27 million $\tau^+\tau^-$ pairs collected by the CLEO II experiment. We search for second class currents in the decay $\tau \to \omega\pi\nu_\tau$ using spin-parity analysis and establish an upper limit on the non-vector current contribution. The mass and width of the $\rho'$ resonance are extracted from a fit to the $\tau \to \omega\pi\nu_\tau$ spectral function. A partial wave analysis of the resonant structure of the $\tau \to 3\pi\pi^0\nu_\tau$ decay is performed; the spectral decomposition of the four pion system is dominated by the $\omega\pi$ and $a_1 \pi$ final states.Comment: 34 pages postscript, also available through http://w4.lns.cornell.edu/public/CLN

Determination of the Michel Parameters and the tau Neutrino Helicity in tau Decay

Using the CLEO II detector at the $e^+e^-$ storage ring CESR, we have determined the Michel parameters $\rho$, $\xi$, and $\delta$ in $\tau^\mp \to l^\mp\nu\bar{\nu}$ decay as well as the tau neutrino helicity parameter $h_{\nu_\tau}$ in $\tau^\mp \to \pi^\mp\pi^0\nu$ decay. From a data sample of $3.02\times 10^6$ tau pairs produced at $\sqrt{s}=10.6 GeV$, using events of the topology $e^+e^- \to \tau^+\tau^- \to (l^\pm\nu\bar{\nu}) (\pi^\mp\pi^0\nu)$ and $e^+e^- \to \tau^+\tau^- \to (\pi^\pm\pi^0\bar{\nu}) (\pi^\mp\pi^0\nu)$, and the determined sign of $h_{\nu_\tau}$, the combined result of the three samples is: $\rho = 0.747\pm 0.010\pm 0.006$, $\xi = 1.007\pm 0.040\pm 0.015$, $\xi\delta = 0.745\pm 0.026\pm 0.009$, and $h_{\nu_\tau} = -0.995\pm 0.010\pm 0.003$. The results are in agreement with the Standard Model V-A interaction.Comment: 18 page postscript file, postscript file also available through http://w4.lns.cornell.edu/public/CLN