Superconductivity at high temperatures is expected in elements with low atomic numbers, based in part on conventional BCS (Bardeen-Cooper-Schrieffer) theory. For example, it has been predicted that when hydrogen is compressed to its dense metallic phase (at pressures exceeding 400 GPa), it will become superconducting with a transition temperature above room temperature. Such pressures are difficult to produce in a laboratory setting, so the predictions are not easily confirmed. Under normal conditions lithium is the lightest metal of all the elements, and may become superconducting at lower pressures; a tentative observation of a superconducting transition in Li has been previously reported. Here we show that Li becomes superconducting at pressures greater than 30 GPa, with a pressure-dependent transition temperature (T(c)) of 20 K at 48 GPa. This is the highest observed T(c) of any element; it confirms the expectation that elements with low atomic numbers will have high transition temperatures, and suggests that metallic hydrogen will have a very high T(c). Our results confirm that the earlier tentative claim of superconductivity in Li was correct.

Folded proteins can exist in multiple conformational substates. Each substate reflects a local minimum on the free-energy landscape with a distinct structure. By using ultrafast 2D-IR vibrational echo chemical-exchange spectroscopy, conformational switching between two well defined substates of a myoglobin mutant is observed on the Ϸ50-ps time scale. The conformational dynamics are directly measured through the growth of cross peaks in the 2D-IR spectra of CO bound to the heme active site. The conformational switching involves motion of the distal histidine/E helix that changes the location of the imidazole side group of the histidine. The exchange between substates changes the frequency of the CO, which is detected by the time dependence of the 2D-IR vibrational echo spectrum. These results demonstrate that interconversion between protein conformational substates can occur on very fast time scales. The implications for larger structural changes that occur on much longer time scales are discussed.multidimensional IR spectroscopy ͉ myoglobin ͉ protein dynamics ͉ protein structural change ͉ ultrafast IR

Ultrafast 2D IR vibrational echo spectroscopy is described and a number of experimental examples are given. Details of the experimental method including the pulse sequence, heterodyne detection, and determination of the absorptive component of the 2D spectrum are outlined. As an initial example, the 2D spectrum of the stretching mode of CO bound to the protein myoglobin (MbCO) is presented. The time dependence of the 2D spectrum of MbCO, which is caused by protein structural evolution, is presented and its relationship to the frequency-frequency correlation function is described and used to make protein structural assignments based on comparisons to molecular dynamics simulations. The 2D vibrational echo experiments on the protein horseradish peroxidase are presented. The time dependence of the 2D spectra of the enzyme in the free form and with a substrate bound at the active site are compared and used to examine the influence of substrate binding on the protein's structural dynamics. The application of 2D vibrational echo spectroscopy to the study of chemical exchange under thermal equilibrium conditions is described. 2D vibrational echo chemical exchange spectroscopy is applied to the study of formation and dissociation of organic solute-solvent complexes and to the isomerization around a carbon-carbon single bond of an ethane derivative.

The ubiquitin system is involved in several basic cellular functions. Ubiquitination is carried out by a cascade of three reactions catalysed by the E1, E2 and E3 enzymes. Among these, the E3 ubiquitin-protein ligases have a pivotal role in determining the specificity of the system by recognizing the target substrates through defined targeting motifs. Although RING finger proteins constitute an important family of E3 ligases, only a few post-transcriptional modifications, including phosphorylation, proline hydroxylation and glycosylation, are known to function as recognition signals for E3. Iron regulatory protein 2 (IRP2), a modulator of iron metabolism, is regulated by iron-induced ubiquitination and degradation. Here we show that the RING finger protein HOIL-1 functions as an E3 ligase for oxidized IRP2, suggesting that oxidation is a specific recognition signal for ubiquitination. The oxidation of IRP2 is generated by haem, which binds to IRP2 in iron-rich cells, and by oxygen, indicating that the iron sensing of IRP2 depends on the synthesis and availability of haem.

Enzyme structural dynamics play a pivotal role in substrate binding and biological function, but the influence of substrate binding on enzyme dynamics has not been examined on fast time scales. In this work, picosecond dynamics of horseradish peroxidase (HRP) isoenzyme C in the free form and when ligated to a variety of small organic molecule substrates is studied by using 2D-IR vibrational echo spectroscopy. Carbon monoxide bound at the heme active site of HRP serves as a spectroscopic marker that is sensitive to the structural dynamics of the protein. In the free form, HRP assumes two distinct spectroscopic conformations that undergo fluctuations on a tens-of-picoseconds time scale. After substrate binding, HRP is locked into a single conformation that exhibits reduced amplitudes and slower time-scale structural dynamics. The decrease in carbon monoxide frequency fluctuations is attributed to reduced dynamic freedom of the distal histidine and the distal arginine, which are key residues in modulating substrate binding affinity. It is suggested that dynamic quenching caused by substrate binding can cause the protein to be locked into a conformation suitable for downstream steps in the enzymatic cycle of HRP.horseradish peroxidase ͉ ultrafast

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Neuroglobin (Ngb), a protein in the globin family, is found in vertebrate brains. It binds oxygen reversibly. Compared with myoglobin (Mb), the amino acid sequence has limited similarity, but key residues around the heme and the classical globin fold are conserved in Ngb. The CO adduct of Ngb displays two CO absorption bands in the IR spectrum, referred to as N 3 (distal histidine in the pocket) and N 0 (distal histidine swung out of the pocket), which have absorption spectra that are almost identical with the Mb mutants L29F and H64V, respectively. The Mb mutants mimic the heme pocket structures of the corresponding Ngb conformers. The equilibrium protein dynamics for the CO adduct of Ngb are investigated by using ultrafast 2D-IR vibrational echo spectroscopy by observing the CO vibration's spectral diffusion (2D-IR spectra time dependence) and comparing the results with those for the Mb mutants. Although the heme pocket structure and the CO FTIR peak positions of Ngb are similar to those of the mutant Mb proteins, the 2D-IR results demonstrate that the fast structural fluctuations of Ngb are significantly slower than those of the mutant Mbs. The results may also provide some insights into the nature of the energy landscape in the vicinity of the folded protein free energy minimum.myoglobin mutants ͉ protein dynamics ͉ energy landscape N euroglobin (Ngb) is a recently discovered family of vertebrate globin proteins. Ngb is expressed predominantly in nerve tissue. It has been hypothesized that Ngb facilitates O 2 diffusion to protect neuronal cells from hypoxia and ischemia (1). Ngb concentration in the brain is fairly low, too low to play the role that myoglobin (Mb) plays in the red muscles. However, because the expression level of Ngb is enhanced under hypoxic conditions in vitro as well as ischemia in vivo (2, 3), Ngb may be involved in neuronal responses to ischemia. A much higher concentration of Ngb is found in the retina. The concentration is high enough to deliver O 2 to mitochondria (4) and may facilitate O 2 diffusion in a manner similar to Mb. Another possibility is that Ngb is involved in redox-coupled sensor regulation for signal transduction in the brain. Ngb binds to the GDP-bound form of the ␣-subunit of heterotrimeric G protein under oxidative stress (5).The 3D structures of Ngb from human and mouse, both having 151 amino acids, have been published (6, 7). Comparisons of Ngb with vertebrate Mb and Hb sequences show only minor similarities at the amino acid level (1), but Ngb features the conserved classical globin fold and contains heme (6). The heme iron in both the ferrous and ferric forms of Ngb is hexacoordinated (8), in contrast to mammalian Mb and Hb, which contain pentacoordinated heme iron. Although hexacoordinated heme has been reported in plant, bacteria, and invertebrate globins, its physiological significance is not yet understood (9). In Ngb, an external gaseous ligand must compete with the sixth ligand, the distal histidine (E7 in helix notation), for binding. Several key residues...

One of the challenges in physical chemistry has been understanding how energy flows in a condensed phase from the microscopic viewpoint. To address this, space-resolved information at the molecular scale is required but has been lacking due to experimental difficulties. We succeeded in the real-time mapping of the vibrational energy flow in a protein with the spatial resolution of a single amino acid residue by combining time-resolved resonance Raman spectroscopy and site-directed single-Trp mutagenesis. Anti-Stokes Raman intensities of the Trp residues at different sites exhibited different temporal evolutions, reflecting propagation of the energy released by the heme group. A classical heat transport model was not able to reproduce the entire experimental data set, showing that we need a molecular-level description to explain the energy flow in a protein. The systematic application of our general methodology to proteins with different structural motifs may provide a greatly increased understanding of the energy flow in proteins.