| 1. |
- Klepsch, Mirjam, 1983-, et al.
(författare)
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Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes
- 2008
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Ingår i: Methods. - : Elsevier BV. - 1095-9130 .- 1046-2023. ; 46:2, s. 48-53
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Tidskriftsartikel (refereegranskat)abstract
- In biological membranes many proteins are organized in complexes. The method of choice for the global analysis of the subunits of these complexes is two-dimensional blue native (2D BN)/SDS–PAGE. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS–PAGE. In the currently available protocols the 1st dimension BN gel lanes get distorted during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, rendering them unsuitable for comparative analysis. We have developed a 2D BN/SDS–PAGE protocol where the 1st dimension BN gel is cast on a GelBond PAG film. Immobilization prevents distortion of BN gel lanes, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. 2D BN/SDS–PAGE with an immobilized 1st dimension was used for the comparative analysis of the cytoplasmic membrane proteomes of Escherichia coli cells overexpressing a membrane protein and to create a 2D BN/SDS–PAGE reference map of the E. coli cytoplasmic membrane proteome with 143 identified proteins from 165 different protein spots.
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| 2. |
- Wagner, Samuel, et al.
(författare)
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Biogenesis of MalF and the MalFGK2 maltose transport complex in Escherichia coli requires YidC
- 2008
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Ingår i: Journal of Biological Chemistry. - 0021-9258 .- 1083-351X. ; :283, s. 17881-17890
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Tidskriftsartikel (refereegranskat)abstract
- The polytopic inner membrane protein MalF is a constituent of the MalFGK2 maltose transport complex in Escherichia coli. We have studied the biogenesis of MalF using a combination of in vivo and in vitro approaches. MalF is targeted via the SRP pathway to the Sec/YidC insertion site. Despite close proximity of nascent MalF to YidC during insertion, YidC is not required for the insertion of MalF into the membrane. However, YidC is required for the stability of MalF and the formation of the MalFGK2 maltose transport complex. Our data indicate that YidC supports the folding of MalF into a stable conformation before it is incorporated into the maltose transport complex.
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| 3. |
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| 4. |
- Wagner, Samuel, et al.
(författare)
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Tuning Escherichia coli for membrane protein overexpression
- 2008
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 105:38, s. 14371-17376
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Tidskriftsartikel (refereegranskat)abstract
- A simple generic method for optimizing membrane protein overexpression in Escherichia coli is still lacking. We have studied the physiological response of the widely used “Walker strains” C41(DE3) and C43(DE3), which are derived from BL21(DE3), to membrane protein overexpression. For unknown reasons, overexpression of many membrane proteins in these strains is hardly toxic, often resulting in high overexpression yields. By using a combination of physiological, proteomic, and genetic techniques we have shown that mutations in the lacUV5 promoter governing expression of T7 RNA polymerase are key to the improved membrane protein overexpression characteristics of the Walker strains. Based on this observation, we have engineered a derivative strain of E. coli BL21(DE3), termed Lemo21(DE3), in which the activity of the T7 RNA polymerase can be precisely controlled by its natural inhibitor T7 lysozyme (T7Lys). Lemo21(DE3) is tunable for membrane protein overexpression and conveniently allows optimizing overexpression of any given membrane protein by using only a single strain rather than a multitude of different strains. The generality and simplicity of our approach make it ideal for high-throughput applications.
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| 5. |
- Abdo, A. A., et al.
(författare)
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FERMI/LARGE AREA TELESCOPE DISCOVERY OF GAMMA-RAY EMISSION FROM THE FLAT-SPECTRUM RADIO QUASAR PKS 1454-354
- 2009
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Ingår i: Astrophysical Journal. - : American Astronomical Society. - 0004-637X .- 1538-4357. ; 697:1, s. 934-941
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Tidskriftsartikel (refereegranskat)abstract
- We report the discovery by the Large Area Telescope (LAT) onboard the Fermi Gamma-Ray Space Telescope of high-energy gamma-ray (GeV) emission from the flat-spectrum radio quasar PKS 1454-354 (z = 1.424). On 2008 September 4, the source rose to a peak flux of (3.5 +/- 0.7) x 10(-6) ph cm(-2) s(-1) (E > 100 MeV) on a timescale of hours and then slowly dropped over the following 2 days. No significant spectral changes occurred during the flare. Fermi/LAT observations also showed that PKS 1454-354 is the most probable counterpart of the unidentified EGRET source 3EG J1500-3509. Multiwavelength measurements performed during the following days (7 September with Swift; 6-7 September with the ground-based optical telescope Automated Telescope for Optical Monitoring; 13 September with the Australia Telescope Compact Array) resulted in radio, optical, UV, and X-ray fluxes greater than archival data, confirming the activity of PKS 1454-354.
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| 6. |
- Abdo, A. A., et al.
(författare)
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MULTIWAVELENGTH MONITORING OF THE ENIGMATIC NARROW-LINE SEYFERT 1 PMN J0948+0022 IN 2009 MARCH-JULY
- 2009
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Ingår i: Astrophysical Journal. - : American Astronomical Society. - 0004-637X .- 1538-4357. ; 707:1, s. 727-737
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Tidskriftsartikel (refereegranskat)abstract
- Following the recent discovery of gamma rays from the radio-loud narrow-line Seyfert 1 galaxy PMN J0948+0022 (z = 0.5846), we started a multiwavelength campaign from radio to gamma rays, which was carried out between the end of 2009 March and the beginning of July. The source displayed activity at all the observed wavelengths: a general decreasing trend from optical to gamma-ray frequencies was followed by an increase of radio emission after less than two months from the peak of the gamma-ray emission. The largest flux change, about a factor of about 4, occurred in the X-ray band. The smallest was at ultraviolet and near-infrared frequencies, where the rate of the detected photons dropped by a factor 1.6-1.9. At optical wavelengths, where the sampling rate was the highest, it was possible to observe day scale variability, with flux variations up to a factor of about 3. The behavior of PMN J0948+0022 observed in this campaign and the calculated power carried out by its jet in the form of protons, electrons, radiation, and magnetic field are quite similar to that of blazars, specifically of flat-spectrum radio quasars. These results confirm the idea that radio-loud narrow-line Seyfert 1 galaxies host relativistic jets with power similar to that of average blazars.
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| 7. |
- Acero, F., et al.
(författare)
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HESS upper limits on very high energy gamma-ray emission from the microquasar GRS 1915+105
- 2009
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Ingår i: Astronomy and Astrophysics. - : EDP Sciences. - 0004-6361 .- 1432-0746. ; 508:3, s. 1135-1140
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Tidskriftsartikel (refereegranskat)abstract
- Context. High energy particles reside in the relativistic jets of microquasars, making them possible sources of very high energy radiation (VHE, > 100 GeV). Detecting this emission would provide a new handle on jet physics. Aims. Observations of the microquasar GRS 1915+105 with the HESS telescope array were undertaken in 2004-2008 to search for VHE emission. Methods. Stereoscopic imaging of Cherenkov radiation from extensive air showers is used to reconstruct the energy and direction of the incident gamma rays. Results. There is no evidence for a VHE gamma-ray signal either from the direction of the microquasar or its vicinity. An upper limit of 6.1 x 10(-13) ph cm(-2) s(-1) (99.9% confidence level) is set on the photon flux above 410 GeV, equivalent to a VHE luminosity of similar to 10(34) erg s(-1) at 11 kpc. Conclusions. The VHE to X-ray luminosity ratio in GRS 1915+105 is at least four orders of magnitude lower than the ratio observed in gamma-ray binaries. The VHE radiative efficiency of the compact jet is less than 0.01% based on its estimated total power of 10(38) erg s(-1). Particle acceleration in GRS 1915+105 is not efficient at high energies and/or the magnetic field is too strong. It is also possible that VHE gamma-rays are produced by GRS 1915+105, but the emission is highly time-dependent.
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| 8. |
- Aharonian, F., et al.
(författare)
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SIMULTANEOUS OBSERVATIONS OF PKS 2155-304 WITH HESS, FERMI, RXTE, AND ATOM : SPECTRAL ENERGY DISTRIBUTIONS AND VARIABILITY IN A LOW STATE
- 2009
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Ingår i: Astrophysical Journal Letters. - 2041-8205 .- 0004-637X .- 1538-4357. ; 696:2, s. L150-L155
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Tidskriftsartikel (refereegranskat)abstract
- We report on the first simultaneous observations that cover the optical, X-ray, and high-energy gamma-ray bands of the BL Lac object PKS 2155-304. The gamma-ray bands were observed for 11 days, between 2008 August 25 and 2008 September 6 (MJD 54704-54715), jointly with the Fermi Gamma-ray Space Telescope and the HESS atmospheric Cherenkov array, providing the first simultaneous MeV-TeV spectral energy distribution (SED) with the new generation of gamma-ray telescopes. The ATOM telescope and the RXTE and Swift observatories provided optical and X-ray coverage of the low-energy component over the same time period. The object was close to the lowest archival X-ray and very high energy (VHE; > 100 GeV) state, whereas the optical flux was much higher. The light curves show relatively little (similar to 30%) variability overall when compared to past flaring episodes, but we find a clear optical/VHE correlation and evidence for a correlation of the X-rays with the high-energy spectral index. Contrary to previous observations in the flaring state, we do not find any correlation between the X-ray and VHE components. Although synchrotron self-Compton models are often invoked to explain the SEDs of BL Lac objects, the most common versions of these models are at odds with the correlated variability we find in the various bands for PKS 2155-304.
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| 9. |
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| 10. |
- Chang-Claude, Jenny, et al.
(författare)
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Age at menarche and menopause and breast cancer risk in the International BRCA1/2 Carrier Cohort Study
- 2007
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Ingår i: Cancer Epidemiology Biomarkers and Prevention. - 1055-9965. ; 16:4, s. 740-746
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Tidskriftsartikel (refereegranskat)abstract
- Background: Early menarche and late menopause are important risk factors for breast cancer, but their effects on breast cancer risk in BRCA1 and BRCA2 carriers are unknown. Methods: We assessed breast cancer risk in a large series of 1,187 BRCA1 and 414 BRCA2 carriers from the International BRCA1/2 Carrier Cohort Study. Rate ratios were estimated using a weighted Cox-regression approach. Results: Breast cancer risk was not significantly related to age at menopause {hazard ratio [HR] for menopause below age 35 years, 0.60 [95% confidence interval (95% CI), 0.25-1.44]; 35 to 40 years, 1.15 [0.65-2.04]; 45 to 54 years, 1.02 [0.65-1.60]; ≥55 years, 1.12 [0.12-5.02], as compared with premenopausal women}. However, there was some suggestion of a reduction in risk after menopause in BRCA2 carriers. There was some evidence of a protective effect of oophorectomy (HR, 0.56; 95% CI, 0.29-1.09) and a significant trend of decreasing risk with increasing time since oophorectomy, but no apparent effect of natural menopause. There was no association between age at menarche and breast cancer risk, nor any apparent association with the estimated total duration of breast mitotic activity. Conclusions: These results are consistent with other observations suggesting a protective effect of oophorectomy, similar in relative effect to that in the general population. The absence of an effect of age at natural menopause is, however, not consistent with findings in the general population and may reflect the different natural history of the disease in carriers.
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