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《Bioorganic & medicinal chemistry》2014,22(13):3316-3324
Desymmetrization of the pseudochiral (2r)-configured cyclohexane-1,2,3-triamines 8 with dimethyl oxalate led to racemic aminoquinoxaline-2,3-diones 9. Selective introduction of the κ pharmacophoric structural elements pyrrolidine and 3,4-dichlorophenylacetamide with a two-carbon distance afforded conformationally restricted κ agonists 13–15 based on the quinoxaline ring system. In competitive radioligand receptor binding studies the benzylamine 13b, the secondary amine 14b, and the carbamate 15 displayed high κ receptor affinity. The Ki value of the lead compound derived methoxycarbonyl derivative 15 is 9.7 nM. However, the κ affinity of 15 is exceeded by 13b and 14b with a basic functional group instead of the methoxycarbonyl group in 1-position of the quinoxaline system. The chlorine atoms of the dichlorophenylacetyl residue are essential, since the corresponding phenylacetyl analogs show considerably reduced κ affinity. The potent κ ligands 13b, 14b and 15 are selective over the related μ- and δ-opioid receptors, σ1, σ2 and NMDA receptors. In the [35S]GTPγS-binding assay 13b behaved as partial agonist with lower activity than U-69,593. 相似文献
4.
RNA polymerase from the archaebacterium Sulfolobus acidocaldarius was chemically modified with AMP o-formylphenyl ester followed by reduction with borohydride. The modified protein catalyzes the labeling of its own largest subunit when incubated with [-33P]UTP in the presence of poly[d(A-T)]. On cleaving of the labeled protein using cyanogen bromide, hydroxylamine or amino acid-specific endoproteinases for a very brief period, the pattern and size of the radioactive fragments formed are best explained by attachment of the label between Gly843 and Met895 of the largest subunit. In this region there exists a highly conserved sequence which is also found in other archaebacterial, eukaryotic and prokaryotic RNA polymerases. This suggests that the binding site for the initiating substrate of RNA polymerases has been conserved during evolution. 相似文献
5.
Mark L. Paddock Scott H. Rongey Edward C. Abresch George Feher Melvin Y. Okamura 《Photosynthesis research》1988,17(1-2):75-96
Many herbicides that inhibit photosynthesis in plants also inhibit photosynthesis in bacteria. We have isolated three mutants of the photosynthetic bacterium Rhodobacter sphaeroides that were selected for increased resistance to the herbicide terbutryne. All three mutants also showed increased resistance to the known electron transfer inhibitor o-phenanthroline. The primary structures of the mutants were determined by recombinant DNA techniques. All mutations were located on the gene coding for the L-subunit resulting in these changes Ile229 Met, Ser223 Pro and Tyr222 Gly. The mutations of Ser223 is analogous to the mutation of Ser264 in the D1 subunit of photosystem II in green plants, strengthening the functional analogy between D1 and the bacterial L-subunit. The changed amino acids of the mutant strains form part of the binding pocket for the secondary quinone, Q
b
. This is consistent with the idea that the herbicides are competitive inhibitors for the Q
b
binding site. The reaction centers of the mutants were characterized with respect to electron transfer rates, inhibition constants of terbutryne and o-phenanthroline, and binding constants of the quinone UQ0 and the inhibitors. By correlating these results with the three-dimensional structure obtained from x-ray analysis by Allen et al. (1987a, 1987b), the likely positions of o-phenanthroline and terbutryne were deduced. These correspond to the positions deduced by Michel et al. (1986a) for Rhodopseudomonas viridis.Abbreviations ATP
adenosine 5-triphosphate
- Bchl
bacteriochlorophyll
- Bphe
bacteriopheophytin
- bp
basepair
- cyt c2+
reduced form of cytochrome c
- DEAE
diethylami-noethyl
- EDTA
ethylenediamine tetraacetic acid
- Fe2+
non-heme iron atom
- LDAO
lauryl dimethylamine oxide
- Pipes
piperazine-N,N-bis-2-ethane-sulfonic acid
- PSII
photosystem II
- RC
reaction center
- SDS
sodium dodecylsulfate
- Tris
tris(hydroxy-methyl)aminomethane
- UQ0
2,3-dimethoxy-5-methyl benzoquinone
- UQ10
ubiquinone 50 相似文献
6.
7.
Rapid and simple isolation of pure photosystem II core and reaction center particles from spinach 总被引:2,自引:2,他引:0
Peter J. van Leeuwen Maaike C. Nieveen Erik Jan van de Meent Jan P. Dekker Hans J. van Gorkom 《Photosynthesis research》1991,28(3):149-153
Pure and active oxygen-evolving PS II core particles containing 35 Chl per reaction center were isolated with 75% yield from spinach PS II membrane fragments by incubation with n-dodecyl--D-maltoside and a rapid one step anion-exchange separation. By Triton X-100 treatment on the column these particles could be converted with 55% yield to pure and active PS II reaction center particles, which contained 6 Chl per reaction center.Abbreviations Bis-Tris
bis[2-hydroxyethyl]imino-tris[hydroxymethyl]methane
- Chl
chlorophyll
- CP29
Chl a/b protein of 29 kDa
- Cyt b
559
cytochrome b
559
- DCBQ
2,5-dichloro-p-benzo-quinone
- LHC II
light-harvesting complex II, predominant Chl a/b protein
- MES
2-[N-Morpholino]ethanesulfonic acid
- Pheo
pheophytin
- PS H
photosystem II
- QA
bound plastoquinone, serving as the secondary electron acceptor in PS II (after Pheo)
- SDS
sodiumdodecylsulfate 相似文献
8.
在麻醉猫,经推动脉注入梭曼、VX,沙林及乙酰甲胆碱引起呼吸中枢严重抑制的剂量分别为0.5—1、3、15、2001μg/头;但在无麻醉、箭毒麻痹、人工呼吸并用药物保护循环的清醒猫,VX用量要增加十多倍,沙林用量增加2~8倍,棱曼用量不变。在严重抑制剂量的给药早期,梭曼使34.8%动物较早地出现膈神经单纤维放电加强,其每次吸气放电的冲动频率由20~30Hz增至50~80Hz,冲动个数由15~25个/每次放电增至40~60个/每次放电,兴奋持续短、迅速转入抑制且不易自动恢复;VX和乙酰甲胆碱使100%动物出现显著的放电加强,其冲动频率由20~30Hz增至70~130Hz、冲动个数由15~25个/每次放电增至60~80个/第次放电,兴奋持续时间较长、转入抑制慢但自动恢复较快;沙林使76.9%动物出现放电加强,其他表现类似VX。三种胆碱酯酶抑制剂和乙酰甲胆碱共使33/52根单纤维放电发生时相变化。结果表明:梭曼对呼吸中枢作用最强、沙林次之、VX最弱且更似乙酰甲胆碱。 相似文献
9.
Richard W. Hendler Barry Bunow John S. Rieske 《Journal of bioenergetics and biomembranes》1985,17(1):51-64
In coenzyme Q-cycles, it is proposed that one electron from the quinol reduces the Rieske iron sulfur center (E
m280 mV) and the remaining electron on the semiquinone reduces cytochromeb
T (E
m–60 mV). TheE
mfor the two-electron oxidation of the quinol is 60 mV and therefore the reduction of cytochromeb
T by quinol is not favorable. As the stability constant for the dismutation of the semiquinone decreases, the calculatedE
mfor the Q/QH couple is lowered to values below theE
mof cytochromeb
T. Contemporary coenzyme Q-cycles are based on the belief that the lower value for theE
mof the Q/QH couple compared to theE
mfor cytochromeb
T means that the semiquinone is a spontaneous reducing agent for theb-cytochrome. The analysis in the paper shows that this is not necessarily so and that neither binding sites nor ionization of the semiquinoneper se alters this situation. For a Q-cycle mechanism to function,ad hoc provisions must be made to drive the otherwise unfavorable reduction of cytochromeb
T by the semiquinone or for the simultaneous transfer of both electrons to cytochromeb
T and cytochromec
1 (or the iron sulfur protein). Q-cycle mechanisms with these additional provisions can explain the observation thus far accumulated. A linear path which is functionally altered by conformational changes may also explain the data. 相似文献
10.
Spectrally pure reaction center preparations from Chloroflexus aurantiacus have been obtained in a stable form; however, the product contained several contaminating polypeptides. The reaction center pigment molecules (probably three bacteriochlorophyll a and three bacteriopheophytin a molecules) are associated with two polypeptides (Mr = 30000 and 28000) in a reaction center complex of Mr = 52000. No carotenoid is present in the complex. These data together with previous spectral data suggest that the Chloroflexus reaction center represents a more primitive evolutionary form of the purple bacterial reaction center, and that it has little if any relationship to the green bacterial component. A reaction center preparation from Rhodopseudomonas sphaeroides R26 was fully denatured at 50°C while the Chloroflexus reaction center required higher temperatures (70–75°C) for complete denaturation. Thus, an intrinsic membrane protein of a photosynthetic thermophile has been demonstrated to have greater thermal stability than the equivalent component of a mesophile. 相似文献