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1.
Cytochrome P-450 isozymes represent a critical component of nature’s spectrum of detoxification catalysts that could be exploited
for bioremediation. The ethanol-inducible human cytochrome P-450 2E1 serves as a model eukaryotic P-450 that complements the
bacterial P-450 cam in dehalogenation and detoxification of environmental pollutants. We explored the construction of novel
chimeric P-450s using cytochrome P-450 camC and 2E1 genes. For construction of chimera 1 (478 amino acids, 55.14 kDa), 145 amino acids from the N-terminus of P-450 2E1 protein
(493 amino acids, 56.84 kDa) were replaced with 130 amino acids from the N-terminus of P-450 camC protein (415 amino acids,
46.66 kDa). In chimera 2 (525 amino acids, 60.24 kDa) the strategy involves replacement of 28 amino acids in the C-terminus
of chimera 1 with 75 amino acids from the C-terminus of P-450 camC gene. Homology models of both the chimeric proteins were
developed using SWISS-MODEL based on the known crystal structure of cytochrome P-450 camC, BM-3, 1DT6A, and 2C17A. The models
indicated that the proposed heme-binding site was intact, which is inevitable for catalytic activity of cytochrome P-450s.
The expression of chimera 1 and 2 genes in Escherichia coli DH5α was evident from light-pink cell pellets, protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis,
and diagnostic carbon monoxide-difference spectra. Our studies show that strategies can be developed to exploit the natural
diversity of the P-450 superfamily to generate chimeric biocatalysts that would provide new templates amenable to directed
evolution. 相似文献
2.
A technique is reported for measuring triplet spin-lattice relaxation times in fluid solution. It is based on the observation of chemically induced electron spin polarization, in the presence of a triplet quencher. Spin-lattice relaxation times of the order of 10 ns are reported for the duroquinone triplet in various solvents. 相似文献
3.
Abazov VM Abbott B Acharya BS Adams M Adams T Alexeev GD Alkhazov G Alton A Alverson G Aoki M Askew A Åsman B Atkins S Atramentov O Augsten K Avila C Badaud F Bagby L Baldin B Bandurin DV Banerjee S Barberis E Baringer P Barreto J Bartlett JF Bassler U Bazterra V Bean A Begalli M Belanger-Champagne C Bellantoni L Beri SB Bernardi G Bernhard R Bertram I Besançon M Beuselinck R Bezzubov VA Bhat PC Bhatia S Bhatnagar V Blazey G Blessing S Bloom K Boehnlein A Boline D Boos EE Borissov G Bose T 《Physical review letters》2012,108(15):151804
We present a measurement of the W boson mass using data corresponding to 4.3 fb(-1) of integrated luminosity collected with the D0 detector during Run II at the Fermilab Tevatron pp collider. With a sample of 1,677,394 W → eν candidate events, we measure M(W) = 80.367 ± 0.026 GeV. This result is combined with an earlier D0 result determined using an independent Run II data sample, corresponding to 1 fb(-1) of integrated luminosity, to yield M(W) = 80.375 ± 0.023 GeV. 相似文献
4.
Abazov VM Abbott B Acharya BS Adams M Adams T Alexeev GD Alkhazov G Alton A Alverson G Alves GA Aoki M Askew A Åsman B Atkins S Atramentov O Augsten K Avila C BackusMayes J Badaud F Bagby L Baldin B Bandurin DV Banerjee S Barberis E Baringer P Barreto J Bartlett JF Bassler U Bazterra V Bean A Begalli M Belanger-Champagne C Bellantoni L Beri SB Bernardi G Bernhard R Bertram I Besançon M Beuselinck R Bezzubov VA Bhat PC Bhatnagar V Blazey G Blessing S Bloom K Boehnlein A Boline D Boos EE 《Physical review letters》2012,108(3):032004
We present a measurement of the ratio of events with correlated t and t ?t spins to the total number of t ?t events. This ratio f is evaluated using a matrix-element-based approach in 729 t ?t candidate events with a single lepton ? (electron or muon) and at least four jets. The analyzed p ?p collisions data correspond to an integrated luminosity of 5.3 fb(-1) and were collected with the D0 detector at the Fermilab Tevatron collider operating at a center-of-mass energy √s=1.96 TeV. Combining this result with a recent measurement of f in dileptonic final states, we find f in agreement with the standard model. In addition, the combination provides evidence for the presence of spin correlation in t ?t events with a significance of more than 3 standard deviations. 相似文献
5.
Ryan Atkins 《Journal of luminescence》2008,128(9):1463-1470
We have completed a study of the optical properties of SrY2O4:Eu3+ under vacuum ultraviolet (VUV) excitation. Reflectance measurements on the undoped material yield a calculated band gap of about 6.1 eV. Studies on the doped material indicate that Eu3+ occupies the Y(1) and Y(2) sites in this host. Host-to-activator energy transfer calculations indicate a preference for transfer to Eu3+ at the Y(2) site. Modeling of the transfer efficiency data leads us to estimate that about 35% of absorbed radiation is lost to the surface under excitation near the band edge. 相似文献
6.
Dabrowski MJ Schrag ML Wienkers LC Atkins WM 《Journal of the American Chemical Society》2002,124(40):11866-11867
Cytochrome P450 monooxygenases (CYPs) metabolize nearly all drugs and toxins. Recently, it has become clear that CYPs exhibit both homotropic and heterotropic allosteric kinetics for many substrates. However, the mechanism of cooperative kinetics has not been established for any specific human CYP/substrate combination. Suggested mechanisms include binding of multiple substrates within distinct, static, subsites of a single large active site or binding of multiple substrates within a single fluid active site. CYP3A4 hydroxylates pyrene with positive cooperativity. Therefore, experiments were designed to exploit the fluorescence properties of pyrene, which diagnostically distinguish between pyrene.pyrene complexes versus spatially separated pyrene substrates. Pyrene complexes (excimers) yield an emission spectrum clearly distinct from pyrene monomers. In lipid-free aqueous/glycerol solutions of CYP3A4, addition of pyrene affords a concentration-dependent low-spin to high-spin conversion of the CYP3A4 heme prosthetic group, indicating occupancy of the active site by pyrene. Under the same conditions, in the presence of CYP3A4 but not other heme proteins, the excimer/monomer ratio (E/M) of pyrene was decreased in emission spectra, compared to pyrene alone. However, excitation spectra indicate a CYP3A4-dependent increase in the wavelength shift for the excimer excitation spectrum versus the monomer excitation spectrum, as well as changes in the excimer excitation peak shape and vibronic structure. These changes are reversed by the CYP3A4 substrate testosterone. Together, the results demonstrate that pyrene.pyrene ground-state complexes occupy the CYP3A4 active site, and they provide the first spectroscopic evidence for substrate complexes within a single fluid active site. Functional implications include the possibility that turnover rate, regioselectivity, and stereoselectivity of the reaction are determined by the substrate.substrate complex rather than individual substrates. 相似文献
7.
Amanda J. Gregory Jonathan P. Atkins Daryl Burdon Michael Elliott 《European Journal of Operational Research》2013
The purpose of this paper is to learn from Complex Adaptive Systems (CAS) theory to inform the development of Problem Structuring Methods (PSMs) both in general and in the specific context of marine management. The focus on marine management is important because it is concerned with a CAS (formed through the interconnection between natural systems, designed systems and social systems) which exemplifies their particularly ‘wicked’ nature. Recognition of this compels us to take seriously the need to develop tools for knowledge elicitation and structuring which meet the demands of CAS. In marine management, chief among those tools is the DPSIR (Drivers – Pressures – State Changes – Impacts – Responses) model and, although widely applied, the extent to which it is appropriate for dealing with the demands of a CAS is questionable. Such questioning is particularly pertinent in the context of the marine environment where there is a need to not only recognise a broad range of stakeholders (a question of boundary critique) but also to manage competing knowledge (economic, local and scientific) and value claims. Hence this paper emphasises how a CAS perspective might add impetus to the development of a critical perspective on DPSIR and PSM theory and practice to promote a more systemic view of decision-making and policy development. 相似文献
8.
Abazov VM Abbott B Acharya BS Adams M Adams T Alexeev GD Alkhazov G Alton A Alverson G Aoki M Askew A Åsman B Atkins S Atramentov O Augsten K Avila C BackusMayes J Badaud F Bagby L Baldin B Bandurin DV Banerjee S Barberis E Baringer P Barreto J Bartlett JF Bassler U Bazterra V Bean A Begalli M Belanger-Champagne C Bellantoni L Beri SB Bernardi G Bernhard R Bertram I Besançon M Beuselinck R Bezzubov VA Bhat PC Bhatia S Bhatnagar V Blazey G Blessing S Bloom K Boehnlein A Boline D Boos EE 《Physical review letters》2012,108(13):131802
We present a search for Kaluza-Klein (KK) particles predicted by models with universal extra dimensions (UED) using a data set corresponding to an integrated luminosity of 7.3 fb(-1), collected by the D0 detector at a pp center-of-mass energy of 1.96 TeV. The decay chain of KK particles can lead to a final state with two muons of the same charge. This signature is used to set a lower limit on the compactification scale of R(-1)>260 GeV in a minimal UED model. 相似文献
9.
Abazov VM Abbott B Acharya BS Adams M Adams T Alexeev GD Alkhazov G Alton A Alverson G Alves GA Aoki M Askew A Åsman B Atkins S Atramentov O Augsten K Avila C BackusMayes J Badaud F Bagby L Baldin B Bandurin DV Banerjee S Barberis E Baringer P Barreto J Bartlett JF Bassler U Bazterra V Bean A Begalli M Belanger-Champagne C Bellantoni L Beri SB Bernardi G Bernhard R Bertram I Besançon M Beuselinck R Bezzubov VA Bhat PC Bhatnagar V Blazey G Blessing S Bloom K Boehnlein A Boline D Boos EE 《Physical review letters》2012,108(18):181803
We study WW and WZ production with ?νqq (?=e,μ) final states using data collected by the D0 detector at the Fermilab Tevatron Collider corresponding to 4.3 fb(-1) of integrated luminosity from pp collisions at sqrt[s]=1.96 TeV. Assuming the ratio between the production cross sections σ(WW) and σ(WZ) as predicted by the standard model, we measure the total WV (V=W,Z) cross section to be σ(WV)=19.6(-3.0)(+3.2) pb and reject the background-only hypothesis at a level of 7.9 standard deviations. We also use b-jet discrimination to separate the WZ component from the dominant WW component. Simultaneously fitting WW and WZ contributions, we measure σ(WW)=15.9(-3.2)(+3.7) pb and σ(WZ)=3.3(-3.3)(+4.1) pb, which is consistent with the standard model predictions. 相似文献
10.
Ram Raghava Robert M. Caddell Lynn Buege Anthony G. Atkins 《Journal of Macromolecular Science: Physics》2013,52(4):655-665
The through thickness total strains of hydrostatically bulged domes of materials are usually determined indirectly. The in-plane circumferential strains are readily measured, and then constancy of volume is invoked to determine the “missing” or unknown strain. Some of our recent work with polymers has shown that volume constancy in large-scale deformation does not hold so that the usual computations of stress and strain during bulging of sheet may be in error. We describe an electrical device that reduces such errors by directly measuring the through thickness (and, thus, the thickness “strain”) for nonmetallic materials. 相似文献