N-Propylnoraporphin-11-O-yl carboxylic esters as potent dopamine D(2) and serotonin 5-HT(1A) receptor dual ligands

A small series of N-propylnoraporphin-11-O-yl carboxylic esters with variant ester lengths were synthesized and their binding potencies at dopamine receptors (D(1), D(2)) and serotonin receptors (5-HT(1A), 5-HT(2A)) were evaluated. Monoesters 3a-f showed binding potency of 100 nM or less for the D(2) receptor, and potency of 10-30 nM for the 5-HT(1A) receptor. Butyryl ester 3d was found to be the best compound possessing the highest potency for both receptors, with K(i) values of 55 and 12 nM for D(2) and 5-HT(1A) receptors, respectively. There is no correlation between the binding potency and the length of the monoesters, but the diesters 9 and 10 were inactive for the D(2) receptor. The dual binding profile of these monoesters for the D(2) and 5-HT(1A) receptors may be useful for the treatment of neuropsychiatric disorders.     

Synthesis and pharmacological investigation of novel 2-aminothiazole-privileged aporphines

A series of apomorphine ((-)-1, APO)-derived analogues ((+/-)-3, (-)-4-(-)-6) were designed and synthesized by hybridizing APO with a privileged 2-aminothiazole functionality which was lent from the orally available anti-parkinsonian drug, pramipexole (2). Among these hybridized compounds, catecholic aporphine (-)-6 shows good affinity at the D(2) receptor with K(i) of 328nM, slightly less potent (3-fold), but more selective against the D(1) receptor than that of the parent compound, APO. Although possessing reduced affinity at the D(2) receptor, aporphines 15 and 18 show significant potency at both the D(1) and 5-HT(1A) receptors. The former compound is equipotent at both receptors (K(i): 116 and 151nM, respectively), while the latter is 8-fold more potent at the D(1) (K(i): 78nM) than at the 5-HT(1A) receptors (K(i): 640nM). These results indicate that the catechol fragment is critical for the D(2) receptor binding of the anti-parkinsonian drug, APO ((-)-1), but not necessary for binding at the D(1) and 5-HT(1A) receptors.     

夜间低温胁迫对两种生长光强下藤黄幼苗叶片荧光特征和活性氧代谢的影响

对两种不同生长光强下（自然光的8％和50％）西双版纳热带雨林木本植物藤黄（Garcinia han-buryi）幼苗经夜间低温（4℃）处理后荧光特性和活性氧代谢的研究结果表明,低温使藤黄叶片光合机构PSⅡ原初光能转化效率（Fv/Fm）、PSⅡ非环式电子传递的量子效率（ФPSⅡ）、非光化学猝灭系数（NPQ）下降,原初荧光（F0）上升。低温胁迫消除后,生长在50％光强下藤黄叶片的Fv/Fm和F0在3d后仍不能完全恢复,而生长在8％光强下藤黄叶片的Fv/Fm和F0基本恢复,说明低温使生长在8％光强下藤黄的光合机构PSⅡ反应中心受到可逆失活,而生长在50％光强下藤黄的光合机构受到氧化伤害。随着低温胁迫时间的延长,两种生长光强藤黄叶片活性氧保护酶（SOD,CAT,APX）的活性虽升高,但O2^-的生成速率、H2O2和MDA含量积累增加。而在恢复阶段,生长在8％光强比生长在50％光强下藤黄叶片的活性氧含量下降得快,进一步说明生长在高光强的植物比生长在低光强的植物受低温伤害大。     

Monitoring enzyme catalysis in the multimeric state: Direct observation of Arthrobacter 4-hydroxybenzoyl-coenzyme A thioesterase catalytic complexes using time-resolved electrospray ionization mass spectrometry

The ability to examine real-time reaction kinetics for multimeric enzymes in their native state may offer unique insights into understanding the catalytic mechanism and its interplay with three-dimensional structure. In this study, we have used a time-resolved electrospray mass spectrometry approach to probe the kinetic mechanism of 4-hydroxybenzoyl-coenzyme A (4-HBA-CoA) thioesterase from Arthrobacter sp. strain SU in the millisecond time domain. Intact tetrameric complexes of 4-HBA-CoA thioesterase with up to four natural substrate (4-HBA-CoA) molecules bound were detected at times as early as 6 ms using an online rapid-mixing device directly coupled to an electrospray ionization time-of-flight mass spectrometer. Species corresponding to the formation of a folded tetramer of the thioesterase at charge states 16+, 17+, 18+, and 19+ around m/z 3800 were observed and assigned as individual tetramers of thioesterase and noncovalent complexes of the tetramers with up to four substrate and/or product molecules. Real-time evaluation of the reaction kinetics was accomplished by monitoring change in peak intensity corresponding to the substrate and product complexes of the tetrameric protein. The mass spectral data suggest that product 4-HBA is released from the active site of the enzyme prior to the release of product CoA following catalytic turnover. This study demonstrates the utility of this technique to provide additional molecular details for an understanding of the individual enzyme states during the thioesterase catalysis and ability to observe real-time interactions between enzyme and substrates and/or products in the millisecond time range.     

Contribution of mobile genetic elements to Desulfovibrio vulgaris genome plasticity

The genome of Desulfovibrio vulgaris strain DePue, a sulfate-reducing Deltaproteobacterium isolated from heavy metal-impacted lake sediment, was completely sequenced and compared with the type strain D. vulgaris Hildenborough. The two genomes share a high degree of relatedness and synteny, but harbour distinct prophage and signatures of past phage encounters. In addition to a highly variable phage contribution, the genome of strain DePue contains a cluster of open-reading frames not found in strain Hildenborough coding for the production and export of a capsule exopolysaccharide, possibly of relevance to heavy metal resistance. Comparative whole-genome microarray analysis on four additional D. vulgaris strains established greater interstrain variation within regions associated with phage insertion and exopolysaccharide biosynthesis.     

异三聚体G蛋白在NAA诱导的拟南芥根生长发育中的作用

以拟南芥的野生型(ws)、异三聚体G蛋白α亚基基因GPA1缺失突变体(gpa1-1,gpa1-2)和超表达突变体(wGα,cGα)为材料,通过施加不同浓度(0~0.2 mg/L)的NAA处理,对拟南芥根生长发育的一些形态指标进行了观测比较.结果表明:(1)随着培养基中NAA浓度的不断升高,5种基因型主根的伸长生长均受到抑制,且抑制作用随浓度升高而增强;4种突变体和野生型主根的生长在相同浓度NAA处理下,无明显差异;(2)NAA在一定浓度范围内,对拟南芥侧根的生长发育起促进作用;在NAA诱导的侧根生长中,G蛋白超表达突变体比野生型更敏感,缺失突变体则不敏感.初步证明G蛋白不参与主根生长发育的调节,而在侧根生长发育中可能起正调节作用.     

Kinetic effect of a downstream strand and its 5'-terminal moieties on single nucleotide gap-filling synthesis catalyzed by human DNA polymerase lambda

During short-patch base excision repair, the excision of a 5'-terminal 2-deoxyribose-5-phosphate moiety of the downstream strand by the 5'-2-deoxyribose-5-phosphate lyase activity of either DNA polymerase beta or lambda is believed to occur after each respective enzyme catalyzes gap-filling DNA synthesis. Yet the effects of this 5'-terminal 2-deoxyribose-5-phosphate moiety on the polymerase activities of these two enzymes have never been quantitatively determined. Moreover, x-ray crystal structures of truncated polymerase lambda have revealed that the downstream strand and its 5'-phosphate group of gapped DNA interact intensely with the dRPase domain, but the kinetic effect of these interactions is unclear. Here, we utilized pre-steady state kinetic methods to systematically investigate the effect of a downstream strand and its 5'-moieties on the polymerase activity of the full-length human polymerase lambda. The downstream strand and its 5'-phosphate were both found to increase nucleotide incorporation efficiency (kp/Kd) by 15 and 11-fold, respectively, with the increase procured by the effect on the nucleotide incorporation rate constant kp rather than the ground state nucleotide binding affinity Kd. With 4 single nucleotide-gapped DNA substrates containing a 1,2-dideoxyribose-5-phosphate moiety, a 2-deoxyribose-5-phosphate mimic, we measured the incorporation efficiencies of 16 possible nucleotides. Our results demonstrate that although this 5'-terminal 2-deoxyribose-5-phosphate mimic does not affect the fidelity of polymerase lambda, it moderately decreased the polymerase efficiency by 3.4-fold. Moreover, this decrease in polymerase efficiency is due to a drop of similar magnitude in kp rather than Kd. The implication of the downstream strand and its 5'-moieties on the kinetics of gap-filling synthesis is discussed.