Development of a high-performance and cost-effective in-vacuum undulator.

In-vacuum undulators (IVUs), which have become an essential tool in synchrotron radiation facilities, have two technical challenges toward further advancement: one is a strong attractive force between top and bottom magnetic arrays, and the other is a stringent requirement on magnetic materials to avoid demagnetization. The former imposes a complicated design on mechanical and vacuum structures, while the latter limits the possibility of using high-performance permanent magnets. To solve these issues, a number of technical developments have been made, such as force cancellation and modularization of magnetic arrays, and enhancement of resistance against demagnetization by means of a special magnetic circuit. The performance of a new IVU built upon these technologies has revealed their effectiveness for constructing high-performance IVUs in a cost-effective manner.

Experimental Demonstration to Control the Pulse Length of Coherent Undulator Radiation by Chirped Microbunching.

In seeded free electron lasers (FELs), the temporal profile of FEL pulses usually reflects that of the seed pulse, and, thus, shorter FEL pulses are available with shorter seed pulses. In an extreme condition, however, this correlation is violated; the FEL pulse is stretched by the so-called slippage effect in undulators, when the seed pulse is ultimately short, e.g., few-cycles long. In a previous Letter, we have proposed a scheme to suppress the slippage effect and reduce the pulse length of FELs ultimately down to a single-cycle duration, which is based on "chirped microbunching," or an electron density modulation with a varying modulation period. Toward realization of FELs based on the proposed scheme, experiments have been carried out to demonstrate its fundamental mechanism in the NewSUBARU synchrotron radiation facility, using an ultrashort seed pulse with the pulse length shorter than five cycles. Experimental results of spectral and cross-correlation measurements have been found to be in reasonable agreement with the theoretical predictions, which strongly suggests the successful demonstration of the proposed scheme.

Development of an undulator with a variable magnetic field profile.

An undulator generating a magnetic field whose longitudinal profile is arbitrarily varied has been developed, which is one of the key components in a number of proposed new concepts in free-electron lasers. The undulator is composed of magnet modules, each of which corresponds to a single undulator period, and is driven by a linear actuator to change the magnetic gap independently. To relax the requirement on the actuator, the mechanical load on each module due to magnetic force acting from opponent and adjacent modules is reduced by means of two kinds of spring systems. The performance of the constructed undulator has been successfully demonstrated by magnetic measurement and characterization of synchrotron radiation.

Transient grating in a thin gas target for characterization of extremely short optical pulses.

A transient-grating cross-correlation frequency-resolved optical gating (TG XFROG) with a thin gas target toward characterization of sub-femtosecond optical pulses is discussed. For evaluation of the reliability, sub-10 fs near-infrared pulses are characterized, the results of which are compared with those given by the sum-frequency-generation XFROG. The TG XFROG covers the nanojoule energy range or that for the advanced few-cycle UV pulses recently reported. It is also shown that the TG XFROG fails to characterize and heavily underestimates the durations of intense test pulses. The FROG technique sensitively detects the onset of this anomalous behavior, which represents a serious issue for pulse characterizations.

Attosecond single-cycle undulator light: a review.

Research at modern light sources continues to improve our knowledge of the natural world, from the subtle workings of life to matter under extreme conditions. Free-electron lasers, for instance, have enabled the characterization of biomolecular structures with sub-ångström spatial resolution, and paved the way to controlling the molecular functions. On the other hand, attosecond temporal resolution is necessary to broaden our scope of the ultrafast world. Here we discuss attosecond pulse generation beyond present capabilities. Furthermore, we review three recently proposed methods of generating attosecond x-ray pulses. These novel methods exploit the coherent radiation of microbunched electrons in undulators and the tailoring of the emitted wavefronts. The computed pulse energy outperforms pre-existing technologies by three orders of magnitude. Specifically, our simulations of the proposed Soft X-ray Laser at MAX IV (Lund, Sweden) show that a pulse duration of 50-100 as and a pulse energy up to 5 [Formula: see text]J is feasible with the novel methods. In addition, the methods feature pulse shape control, enable the incorporation of orbital angular momentum, and can be used in combination with modern compact free-electron laser setups.

Interaction mapping of the Sec61 translocon identifies two Sec61α regions interacting with hydrophobic segments in translocating chains.

Many proteins in organelles of the secretory pathway, as well as secretory proteins, are translocated across and inserted into the endoplasmic reticulum membrane by the Sec61 translocon, a protein-conducting channel. The channel consists of 10 transmembrane (TM) segments of the Sec61α subunit and possesses an opening between TM2b and TM7, termed the lateral gate. Structural and biochemical analyses of complexes of Sec61 and its ortholog SecY have revealed that the lateral gate is the exit for signal sequences and TM segments of translocating polypeptides to the lipid bilayer and also involved in the recognition of such hydrophobic sequences. Moreover, even marginally hydrophobic (mH) segments insufficient for membrane integration can be transiently stalled in surrounding Sec61α regions and cross-linked to them, but how the Sec61 translocon accommodates these mH segments remains unclear. Here, we used Cys-scanned variants of human Sec61α expressed in cultured 293-H cells to examine which channel regions associate with mH segments. A TM segment in a ribosome-associated polypeptide was mainly cross-linked to positions at the lateral gate, whereas an mH segment in a nascent chain was cross-linked to the Sec61α pore-interior positions at TM5 and TM10, as well as the lateral gate. Of note, cross-linking at position 180 in TM5 of Sec61α was reduced by an I179A substitution. We therefore conclude that at least two Sec61α regions, the lateral gate and the pore-interior site around TM5, interact with mH segments and are involved in accommodating them.

Mode-locked laser with a repetition rate of 17.6 THz.

In 2001, a concept was proposed to generate mode-locked ultrashort laser pulses with a repetition rate in excess of 10 THz [Phys. Rev. Lett.87, 223901 (2001)PRLTAO0031-900710.1103/PhysRevLett.87.223901], which has not been demonstrated so far. In the present research, the concept is experimentally demonstrated using a dispersion-compensated high-finesse cavity filled with hydrogen gas. Second-order intensity autocorrelation is used for distinguishing two cases with and without mode-locking as well as for characterization of the temporal profile. Mode-locked sub-30-fs pulses with a repetition rate of 17.6 THz are synthesized by continuous-wave laser lines generated via stimulated Raman scattering and four-wave Raman mixing induced in the high-finesse cavity.

Radiation-induced magnetization reversal causing a large flux loss in undulator permanent magnets.

We report an unexpectedly large flux loss observed in permanent magnets in one of the undulators operated in SACLA, the x-ray free electron laser facility in Japan. Characterizations of individual magnets extracted from the relevant undulator have revealed that the flux loss was caused by a homogeneous magnetization reversal extending over a wide area, but not by demagnetization of individual magnets damaged by radiation. We show that the estimated flux-loss rate is much higher than what is reported in previous papers, and its distribution is much more localized to the upstream side. Results of numerical and experimental studies carried out to validate the magnetization reversal and quantify the flux loss are presented, together with possible countermeasures against rapid degradation of the undulator performance.

Using irregularly spaced current peaks to generate an isolated attosecond X-ray pulse in free-electron lasers.

A method is proposed to generate an isolated attosecond X-ray pulse in free-electron lasers, using irregularly spaced current peaks induced in an electron beam through interaction with an intense short-pulse optical laser. In comparison with a similar scheme proposed in a previous paper, the irregular arrangement of current peaks significantly improves the contrast between the main and satellite pulses, enhances the attainable peak power and simplifies the accelerator layout. Three different methods are proposed for this purpose and achievable performances are computed under realistic conditions. Numerical simulations carried out with the best configuration show that an isolated 7.7 keV X-ray pulse with a peak power of 1.7 TW and pulse length of 70 as can be generated. In this particular example, the contrast is improved by two orders of magnitude and the peak power is enhanced by a factor of three, when compared with the previous scheme.

Stability and flexibility of marginally hydrophobic-segment stalling at the endoplasmic reticulum translocon.

Many membrane proteins are integrated into the endoplasmic reticulum membrane through the protein-conducting channel, the translocon. Transmembrane segments with insufficient hydrophobicity for membrane integration are frequently found in multispanning membrane proteins, and such marginally hydrophobic (mH) segments should be accommodated, at least transiently, at the membrane. Here we investigated how mH-segments stall at the membrane and their stability. Our findings show that mH-segments can be retained at the membrane without moving into the lipid phase and that such segments flank Sec61α, the core channel of the translocon, in the translational intermediate state. The mH-segments are gradually transferred from the Sec61 channel to the lipid environment in a hydrophobicity-dependent manner, and this lateral movement may be affected by the ribosome. In addition, stalling mH-segments allow for insertion of the following transmembrane segment, forming an Ncytosol/Clumen orientation, suggesting that mH-segments can move laterally to accommodate the next transmembrane segment. These findings suggest that mH-segments may be accommodated at the ER membrane with lateral fluctuation between the Sec61 channel and the lipid phase.

The N-terminal motif of PMP70 suppresses cotranslational targeting to the endoplasmic reticulum.

Many membrane proteins possessing hydrophobic transmembrane (TM) segments are cotranslationally integrated into the endoplasmic reticulum (ER) membrane. Various peroxisomal and mitochondrial membrane proteins escape the ER-targeting mechanism and are targeted to their destinations. Here, we discovered a short segment in the 70-kDa peroxisomal membrane protein (PMP70) that suppresses ER targeting. The first TM segment has an intrinsic signal function that targets the nascent chain to the ER. The ER targeting was suppressed by a short N-terminal sequence of nine residues that is 80 residues upstream of the TM segment. Among the nine residues, Ser(5) is indispensable. The short segment also suppressed the signal peptide function of an authentic secretory protein. This function of the short segment was suppressed by the recombinant motif-GST fusion protein. The 50-kDa and 20-kDa proteins were crosslinked with the motif. The PMP70 molecule with the Ser5Ala point mutation predominantly localized to the ER. We propose the concept of an ER-targeting suppressor that suppresses the ER-targeting mechanism via a binding factor.

Generation of intense subcycle optical pulses in a gas.

The generation of intense subcycle laser pulses during the propagation of two-color femtosecond pulses in a gas medium is investigated theoretically and experimentally. Four-wave mixing induced by the laser pulses in a gas medium generates multi-octave laser radiation from the ultraviolet to the infrared, which forms stable subcycle laser pulses after a certain propagation distance in a gas medium with group-velocity dispersion. The intense subcycle laser pulses would allow the coherent control of the waveforms of soft-x-rays generated via high-harmonic generation.

Resonant and nonresonant multiphoton ionization processes in the mass spectrometry of explosives.

Multiphoton ionization processes were studied for three types of explosives using a line-tunable ultraviolet femtosecond laser. When peroxides such as triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) were ionized through a nonresonant two-photon process, a molecular ion was dominantly observed by reducing the excess energy remaining in the ion. However, an aromatic nitro compound such as 2,4,6-trinitrotoluene (TNT) produced large signals arising from molecular and fragment ions by resonant two-photon ionization. In addition, only fragment ions were produced from a nonaromatic nitro compound such as 1,3,5-trinitroperhydro-1,3,5-triazine (RDX), even when a resonant two-photon ionization process was employed, suggesting that a further reduction in excess energy would be necessary if a molecular ion were to be observed.

A few positively charged residues slow movement of a polypeptide chain across the endoplasmic reticulum membrane.

Many polypeptide chains are translocated across and integrated into the endoplasmic reticulum membrane through protein-conducting channels. During the process, amino acid sequences of translocating polypeptide chains are scanned by the channels and classified to be retained in the membrane or translocated into the lumen. We established an experimental system with which the kinetic effect of each amino acid residue on the polypeptide chain movement can be analyzed with a time resolution of tens of seconds. Positive charges greatly slow movement; only two lysine residues caused a remarkable slow down, and their effects were additive. The lysine residue was more effective than arginine. In contrast, clusters comprising three residues of each of the other 18 amino acids had little effect on chain movement. We also demonstrated that a four lysine cluster can exert the effect after being fully exposed from the ribosome. We concluded that as few as two to three residues of positively charged amino acids can slow the movement of the nascent polypeptide chain across the endoplasmic reticulum membrane. This effect provides a fundamental basis of the topogenic function of positively charged amino acids.

Frequency-resolved optical gating with two nonlinear optical processes.

A frequency-resolved optical gating (FROG) technique that combines autocorrelation (second-harmonic-generation FROG, SHG-FROG) and cross correlation FROG (XFROG) is reported for simultaneous characterization of two unknown optical pulses. Two SHG-FROG signals and a XFROG signal are acquired in a single measurement using a single diagnostic system. Unlike the conventional combination of SHG-FROG for reference-pulse characterization and XFROG for test-pulse characterization, the ambiguity in the direction of time in SHG-FROG is removed during phase retrieval by simultaneously analyzing the three FROG signals. Furthermore, overall characterization is faster, more robust, and highly convergent than the conventional combination of SHG-FROG and XFROG techniques.

Membrane translocation of lumenal domains of membrane proteins powered by downstream transmembrane sequences.

Translocation of the N-terminus of a type I signal anchor (SA-I) sequence across the endoplasmic reticulum membrane can be arrested by tagging with a streptavidin-binding peptide tag (SBP tag) and trapping by streptavidin. In the present study, we first examine the affinity required for the translocation arrest. When the SBP tag is serially truncated, the ability for arrest gradually decreases. Surface plasmon resonance analysis shows that an interaction as strong as 10(-8) M or a smaller dissociation constant is required for trapping the topogenesis of a natural SA-I sequence. Such truncated tags, however, become effective by mutating the SA-I sequence, suggesting that the translocation motivation is considerably influenced by the properties of the SA-I sequence. In addition, we introduce the SBP tag into lumenal loops of a multispanning membrane protein, human erythrocyte band 3. Among the tagged loops between transmembrane 1 (TM1) and TM8, three loops are trapped by cytosolic streptavidin. These loops are followed by TM sequences possessing topogenic properties, like the SA-I sequence, and translocation of one loop is diminished by insertion of a proline into the following TM sequence. These findings suggest that the translocation of lumenal loops by SA-I-like TM sequences has a crucial role in topogenesis of multispanning membrane proteins.

Stop-and-move of a marginally hydrophobic segment translocating across the endoplasmic reticulum membrane.

Many membrane proteins are cotranslationally integrated into the endoplasmic reticulum membrane via the protein-conducting channel, the so-called translocon. The hydrophobic transmembrane segment of the translocating nascent polypeptide chain stops at the translocon and then moves laterally into the membrane. Partitioning of the hydrophobic segment into the membrane is the primary determinant for membrane insertion. Here, we examined the behavior of a marginally hydrophobic segment at the translocon and found that its stop-translocation was greatly affected by the C-terminally attached ribosomes. The marginally hydrophobic segment first stops at the membrane and then moves into the lumen as long as the nascent chain is attached to translating ribosomes. When it is released from the ribosome by the termination codon, the marginally hydrophobic segment does not move. Puromycin or RNase treatment also suppressed movement. The movement was reversibly inhibited by high-salt conditions and irreversibly inhibited by ethylenediaminetetraacetic acid. There is an unstable state prior to the stable membrane insertion of the transmembrane segment. This characteristic state is maintained by the synthesizing ribosome.

Phosphatidylserine-binding protein lactadherin inhibits protein translocation across the ER membrane.

Secretory and membrane proteins are translocated across and inserted into the endoplasmic reticulum membrane via translocon channels. To investigate the effect of the negatively-charged phospholipid phosphatidylserine on the translocation of nascent polypeptide chains through the translocon, we used the phosphatidylserine-binding protein lactadherin C2-domain. Lactadherin inhibited targeting of nascent chain to the translocon by signal sequence and the initiation of translocation. Moreover, lactadherin inhibited the movement of the translocating polypeptide chain regardless of the presence or absence of positively-charged residues. Phosphatidylserine might be critically involved in translocon function, but it is not a major determinant for translocation arrest of positively-charged residues.

Tail-extension following the termination codon is critical for release of the nascent chain from membrane-bound ribosomes in a reticulocyte lysate cell-free system.

Nascent chain release from membrane-bound ribosomes by the termination codon was investigated using a cell-free translation system from rabbit supplemented with rough microsomal membrane vesicles. Chain release was extremely slow when mRNA ended with only the termination codon. Tail extension after the termination codon enhanced the release of the nascent chain. Release reached plateau levels with tail extension of 10 bases. This requirement was observed with all termination codons: TAA, TGA and TAG. Rapid release was also achieved by puromycin even in the absence of the extension. Efficient translation termination cannot be achieved in the presence of only a termination codon on the mRNA. Tail extension might be required for correct positioning of the termination codon in the ribosome and/or efficient recognition by release factors.

Sub-10-fs deep-ultraviolet light source with stable power and spectrum.

In some applications of ultrafast spectroscopy that employ sub-10-fs pulses, the pulse spectrum and power need to be stable for several tens of minutes. In this study, we generate sub-10-fs deep-ultraviolet (DUV) pulses with such stabilities by chirped-pulse four-wave mixing. A power fluctuation of less than 3% rms was realized by employing stabilization schemes that employ a power stabilizer. The pulses generated in this study have been applied to transient absorption spectroscopy in the DUV with a sub-10-fs time resolution [Phys. Chem. Chem. Phys.14, 6200 (2012).10.1039/c2cp23649d]. This sub-10-fs DUV source has a similar performance to widely used noncollinear optical parametric amplifiers.