Prospects and challenges for squeezing-enhanced optical atomic clocks.

Optical atomic clocks are a driving force for precision measurements due to the high accuracy and stability demonstrated in recent years. While further improvements to the stability have been envisioned by using entangled atoms, squeezing the quantum mechanical projection noise, evaluating the overall gain must incorporate essential features of an atomic clock. Here, we investigate the benefits of spin squeezed states for clocks operated with typical Brownian frequency noise-limited laser sources. Based on an analytic model of the closed servo-loop of an optical atomic clock, we report here quantitative predictions on the optimal clock stability for a given dead time and laser noise. Our analytic predictions are in good agreement with numerical simulations of the closed servo-loop. We find that for usual cyclic Ramsey interrogation of single atomic ensembles with dead time, even with the current most stable lasers spin squeezing can only improve the clock stability for ensembles below a critical atom number of about one thousand in an optical Sr lattice clock. Even with a future improvement of the laser performance by one order of magnitude the critical atom number still remains below 100,000. In contrast, clocks based on smaller, non-scalable ensembles, such as ion clocks, can already benefit from squeezed states with current clock lasers.

Motional Fock states for quantum-enhanced amplitude and phase measurements with trapped ions.

The quantum noise of the vacuum limits the achievable sensitivity of quantum sensors. In non-classical measurement schemes the noise can be reduced to overcome this limitation. However, schemes based on squeezed or Schrödinger cat states require alignment of the relative phase between the measured interaction and the non-classical quantum state. Here we present two measurement schemes on a trapped ion prepared in a motional Fock state for displacement and frequency metrology that are insensitive to this phase. The achieved statistical uncertainty is below the standard quantum limit set by quantum vacuum fluctuations, enabling applications in spectroscopy and mass measurements.

A mechanism to prevent production of reactive oxygen species by Escherichia coli respiratory complex I.

Respiratory complex I plays a central role in cellular energy metabolism coupling NADH oxidation to proton translocation. In humans its dysfunction is associated with degenerative diseases. Here we report the structure of the electron input part of Aquifex aeolicus complex I at up to 1.8 Å resolution with bound substrates in the reduced and oxidized states. The redox states differ by the flip of a peptide bond close to the NADH binding site. The orientation of this peptide bond is determined by the reduction state of the nearby [Fe-S] cluster N1a. Fixation of the peptide bond by site-directed mutagenesis led to an inactivation of electron transfer and a decreased reactive oxygen species (ROS) production. We suggest the redox-gated peptide flip to represent a previously unrecognized molecular switch synchronizing NADH oxidation in response to the redox state of the complex as part of an intramolecular feed-back mechanism to prevent ROS production.

Reactive Oxygen Species Production by Escherichia coli Respiratory Complex I.

Respiratory complex I couples the electron transfer exclusively from NADH to a quinone with the translocation of protons across the membrane. However, Escherichia coli adapts to imposed high cellular NADPH concentrations by selecting the mutations E183A(F) and E183G(F) that lead to a high catalytic efficiency of complex I with NADPH. Other mutations at position E183(F) resulting in an efficient NADPH oxidation were not selected. Here we show that the naturally occurring variants exhibit a remarkably low level of production of reactive oxygen species, a byproduct of NAD(P)H oxidation, that besides high catalytic efficiency might be favored by natural selection.

Inhibition of Escherichia coli respiratory complex I by Zn(2+).

The energy-converting NADH:ubiquinone oxidoreductase, respiratory complex I, couples NADH oxidation and quinone reduction with the translocation of protons across the membrane. Complex I exhibits a unique L shape with a peripheral arm extending in the aqueous phase and a membrane arm embedded in the lipid bilayer. Both arms have a length of ∼180 Å. The electron transfer reaction is catalyzed by a series of cofactors in the peripheral arm, while the membrane arm catalyzes proton translocation. We used the inhibition of complex I by zinc to shed light on the coupling of the two processes, which is not yet understood. Enzyme kinetics revealed the presence of two high-affinity binding sites for Zn(2+) that are attributed to the proton translocation pathways in the membrane arm. Electrochemically induced Fourier transform infrared difference spectroscopy demonstrated that zinc binding involves at least two protonated acidic residues. Electron paramagnetic resonance spectroscopy showed that one of the cofactors is only partially reduced by NADH in the presence of Zn(2+). We conclude that blocking the proton channels in the membrane arm leads to a partial block of the electron transfer in the peripheral arm, indicating the long-range coupling between both processes.

Engineering the respiratory complex I to energy-converting NADPH:ubiquinone oxidoreductase.

The respiratory complex I couples the electron transfer from NADH to ubiquinone with a translocation of protons across the membrane. Its nucleotide-binding site is made up of a unique Rossmann fold to accommodate the binding of the substrate NADH and of the primary electron acceptor flavin mononucleotide. Binding of NADH includes interactions of the hydroxyl groups of the adenosine ribose with a conserved glutamic acid residue. Structural analysis revealed that due to steric hindrance and electrostatic repulsion, this residue most likely prevents the binding of NADPH, which is a poor substrate of the complex. We produced several variants with mutations at this position exhibiting up to 200-fold enhanced catalytic efficiency with NADPH. The reaction of the variants with NAD(P)H is coupled with proton translocation in an inhibitor-sensitive manner. Thus, we have created an energy-converting NADPH:ubiquinone oxidoreductase, an activity so far not found in nature. Remarkably, the oxidation of NAD(P)H by the variants leads to an enhanced production of reactive oxygen species.

Role of subunit NuoL for proton translocation by respiratory complex I.

The NADH:ubiquinone oxidoreductase, respiratory complex I, couples the transfer of electrons from NADH to ubiquinone with a translocation of protons across the membrane. The complex consists of a peripheral arm catalyzing the electron transfer reaction and a membrane arm involved in proton translocation. The recently published X-ray structures of the complex revealed the presence of a unique 110 Å "horizontal" helix aligning the membrane arm. On the basis of this finding, it was proposed that the energy released by the redox reaction is transmitted to the membrane arm via a conformational change in the horizontal helix. The helix corresponds to the C-terminal part of the most distal subunit NuoL. To investigate its role in proton translocation, we characterized the electron transfer and proton translocation activity of complex I variants lacking either NuoL or parts of the C-terminal domain. Our data suggest that the H+/2e- stoichiometry of the ΔNuoL variant is 2, indicating a different stoichiometry for proton translocation as proposed from structural data. In addition, the same H+/e- stoichiometry is obtained with the variant lacking the C-terminal transmembraneous helix of NuoL, indicating its role in energy transmission.