An unanticipated architecture of the 750-kDa α6β6 holoenzyme of 3-methylcrotonyl-CoA carboxylase

3-Methylcrotonyl-CoA carboxylase (MCC), a member of the biotin-dependent carboxylase superfamily, is essential for the metabolism of leucine, and deficient mutations in this enzyme are linked to methylcrotonylglycinuria (MCG) and other serious diseases in humans. MCC has strong sequence conservation with propionyl-CoA carboxylase (PCC), and their holoenzymes are both 750-kilodalton (kDa) α(6)β(6) dodecamers. Therefore the architecture of the MCC holoenzyme is expected to be highly similar to that of PCC. Here we report the crystal structures of the Pseudomonas aeruginosa MCC (PaMCC) holoenzyme, alone and in complex with coenzyme A. Surprisingly, the structures show that the architecture and overall shape of PaMCC are markedly different when compared to PCC. The α-subunits show trimeric association in the PaMCC holoenzyme, whereas they have no contacts with each other in PCC. Moreover, the positions of the two domains in the β-subunit of PaMCC are swapped relative to those in PCC. This structural information establishes a foundation for understanding the disease-causing mutations of MCC and provides new insights into the catalytic mechanism and evolution of biotin-dependent carboxylases. The large structural differences between MCC and PCC also have general implications for the relationship between sequence conservation and structural similarity.     

矮杨梅叶的抑菌作用

本文采用琼脂扩散法和生物学数理统计,从定量的角度,对贵州产矮杨梅（ＭｙｒｉｃａｎａｎａＣｅｖａｌ）５种变异类型叶的抑菌作用进行了比较研究。结果表明,各类型叶的提取液对Ｇ＋金黄色葡萄球菌,枯草杆菌有抑菌作用,对Ｇ 大肠杆菌,志贺氏痢疾杆菌无抑菌作用。矮杨梅植物种内的抑菌作用存在极显著性差异。     

Fluorescence ratio imaging of cyclic AMP in single cells.

Fluorescence imaging is perhaps the most powerful technique currently available for continuously observing the dynamic intracellular biochemistry of single living cells. However, fluorescent indicator dyes have been available only for simple inorganic ions such as Ca2+, H+, Na+, K+, Mg2+ and Cl-. We now report a fluorescent indicator for the adenosine 3',5'-cyclic monophosphate (cAMP) signalling pathway. The sensor consists of cAMP-dependent protein kinase in which the catalytic (C) and regulatory (R) subunits are each labelled with a different fluorescent dye such as fluorescein or rhodamine capable of fluorescence resonance energy transfer in the holoenzyme complex R2C2. When cAMP molecules bind to the R subunits, the C subunits dissociate, thereby eliminating energy transfer. The change in shape of the fluorescence emission spectrum allows cAMP concentrations and the activation of the kinase to be nondestructively visualized in single living cells microinjected with the labelled holoenzyme.     

大连驯化生物膜对水体铅吸附的研究

通过对比不同浓度铅离子存在条件下生物膜菌群的生长量,判断其对铅离子的耐受性;挑选吸附性强的菌群扩大培养,进一步作菌群驯化和铅的吸附试验;通过驯化前后吸附曲线和膜菌群的镜检观察对比,了解生物膜菌群在铅污染过程中的自净机制,为驯化生物膜处理含铅污染水体作了基础研究.     

驯化生物膜对水体铅吸附的研究

通过对比不同浓度铅离子存在条件下生物膜菌群的生长量,判断其对铅离子的耐受性;挑选吸附性强的菌群扩大培养,进一步作菌群驯化和铅的吸附试验;通过驯化前后吸附曲线和膜菌群的镜检观察对比,了解生物膜菌群在铅污染过程中的自净机制,为驯化生物膜处理含铅污染水体作了基础研究.     

煤炭微生物絮凝剂的研究

微生物絮凝剂属于新型天然高分子絮凝剂,它与传统化工类絮凝剂相比具有絮凝效果显著及环保等优点。本文把酱油曲霉作为煤炭微生物絮凝剂的产生菌,采用离心沉降和超声波破碎等方法,研究了酱油曲霉的培养液原液、培养原液离心上清液、破碎液、破碎液离心上清液等含有不同成分的四种菌体提取液对煤泥水的絮凝效果。絮凝试验结果表明:酱油曲霉具有不同成分的菌体产生液对煤泥水都具有一定的絮凝作用。其中采用破碎液离心上清液,当菌液量为1.5%,在添加助凝剂(CaCl2)用量为0.2g/L的条件下。絮凝率最高可达到90.76%。作为煤炭生物絮凝剂酱油曲霉具有优良的絮凝效果。     

Correction of cell-cell communication defect by introduction of a protein kinase into mutant cells

The cell-to-cell permeability of the junctions of various cultured mammalian cell types depends on the concentration of intracellular cyclic AMP [( cAMP]i). The permeability rises when the cells are supplied with exogenous cyclic AMP or when their cyclic AMP synthesis is stimulated with choleragen or hormones; it falls when [cAMP]i is lowered by application of serum or due to increase in cell density. The rise and fall in permeability take several hours to develop (the rise is protein synthesis-dependent) and they occur concurrently with the rise and fall in the number of intramembrane particles of the gap junctions, which probably embody the cell-to-cell channels. Is this permeability regulation mediated by phosphorylating protein kinase? In many eukaryotes, the cyclic AMP receptor is a protein kinase consisting of a pair of regulatory subunits and a pair of catalytic subunits. The latter dissociate from the holoenzyme as the cyclic AMP binds to the regulatory subunits and, in this dissociated form, catalyse the phosphorylation of the target. The regulatory subunit occurs in two isoenzyme forms, I and II. The catalytic subunit seems invariant; subunits from different isoenzymes can substitute for each other. We show here that a mutant cell lacking the isoenzyme I is deficient in permeable junctions, and that this junctional defect is corrected when the mutant is supplied with exogenous catalytic subunit.     

抗结核分枝杆菌感染的免疫机制

对宿主利用体内各种细胞与结核分支杆菌的相互作用进行了综述.机体抵抗结核分枝杆菌感染的机制包括特异性免疫和非特异性免疫,参与非特异性免疫的主要有巨噬细胞和γδT细胞,另外,树突状细胞在引发T细胞免疫方面起着关键的作用.由于结核分支杆菌是胞内寄生菌,特异性免疫以细胞免疫为主,主要包括CD4+T细胞免疫和CD8+T细胞免疫.     

亚营养对谷氨酸发酵过程中副产物的影响

研究了谷氨酸菌亚营养状态代谢的过程,探讨不同葡萄糖浓度下谷氨酸菌代谢主产物和副产物的变化规律,通过Uniform Design Version软件分析,得到葡萄糖浓度与副产物乳酸的发酵模型.     

两类微生物SOD的比较研究

报道了酵母ＳＯＤ工程菌ＺＨ－１／ｐＳＨ１比耐高温细菌Ｂ．Ｓ２１１－１５的抗Ｈ２Ｏ２水平高，其单位体积培养液和酵得率约为细菌的二倍，但单位细胞酶的含量比细菌低，高温细菌的ＳＯＤ热稳定性和存放稳定比酵母工程菌好，二者在ｐＨ４－１０范围内都很稳定，但对蛋白的耐受性差异很大，其中酵母ＳＯＤ工程勤务产好。     

Crystal structure of a protein phosphatase 2A heterotrimeric holoenzyme

Protein phosphatase 2A (PP2A) is a principal Ser/Thr phosphatase, the deregulation of which is associated with multiple human cancers, Alzheimer's disease and increased susceptibility to pathogen infections. How PP2A is structurally organized and functionally regulated remains unclear. Here we report the crystal structure of an AB'C heterotrimeric PP2A holoenzyme. The structure reveals that the HEAT repeats of the scaffold A subunit form a horseshoe-shaped fold, holding the catalytic C and regulatory B' subunits together on the same side. The regulatory B' subunit forms pseudo-HEAT repeats and interacts with the C subunit near the active site, thereby defining substrate specificity. The methylated carboxy-terminal tail of the C subunit interacts with a highly negatively charged region at the interface between A and B' subunits, suggesting that the C-terminal carboxyl methylation of the C subunit promotes B' subunit recruitment by neutralizing charge repulsion. Together, our structural results establish a crucial foundation for understanding PP2A assembly, substrate recruitment and regulation.     

枯草杆菌GMS7突变株肌苷发酵研究

以肌苷产生菌枯草杆菌7171—9—1为出发菌株,经物理、化学诱变剂连续处理,获得一株腺嘌呤营养缺陷型并对8—氮杂鸟嘌呤、磺胺胍、链霉素有三重抗性的突变株GMS7,在最佳摇瓶发酵条件下,平均产肌苷为21.35g/L,最高可达23.21g/L,比原菌提高了1.4倍。此外从形态学角度,跟踪观测了肌苷发酵过程中菌体形态的变化,并探讨了各种因素对发酵产苷的影响。     

鲢鱼白皮病致病菌特性的研究

对自然发生白皮病的病（死）鲢鱼进行了病原菌的分离与特性鉴定。从６尾新鲜病（死）鲢鱼的病变部位组织分离到几乎纯一且多量的细菌,从每尾鱼的分离物做纯培养３株共１８株,经形态特征检查及理化特性鉴定初步表明为气单胞菌属（Ａｅｒｏｍｏｎａｓ）的细菌,其中５尾为单一的豚鼠气单胞菌（Ａ．ｃａｖｉａｅ）感染,１尾为豚鼠气单胞菌（Ａ．ｃａｖｉａｅ）与嗜水气单胞菌（Ａ．ｈｙｄｒｏｐｈｉｌａ）两种细菌感染。经致病作用检验初步表明所分离到的两种气单胞菌均可构成鲢鱼白皮病的病原菌。经用１５种常用抗菌类药物做药敏试验,结果表明１８株供试菌对链霉素等１２种均高敏,对青霉素等３种均耐药。     

浸矿菌液/天然黄铁矿界面的动态氧化研究

细菌氧化黄铁矿的过程是细菌破坏黄铁矿化学键和建立新化学键的过程·细菌在浸矿菌液/天然黄铁矿界面上产生一系列的生物化学反应·由于黄铁矿的(100),(110),(111)面上亚铁离子和对硫离子团的分布形式、密度不相同,因此菌液/黄铁矿各种界面的氧化腐蚀程度也有差异·通过对天然黄铁矿(100),(110)和(111)面的细菌氧化动态试验研究,可以将细菌氧化黄铁矿的过程分为三个阶段:第一阶段是黄铁矿的点状氧化阶段;第二阶段为黄铁矿面状氧化阶段;第三阶段为黄铁矿遭到严重的氧化腐蚀,最终导致晶体破坏·细菌对黄铁矿不同面氧化强度顺序为(111)>(110)>(100)·     

16S rDNA序列技术分析鉴定颗粒污泥中的微生物

采用16S rDNA序列分析技术对降解五氯酚的微氧颗粒污泥形成过程中真细菌和古细菌的种群多样性和动态变化进行了研究.通过对DGGE主要条带进行序列比对,发现颗粒污泥中真细菌和古细菌都与不可培养的微生物具有很高的相似性,微氧颗粒污泥中同时存在好氧菌、微氧菌和厌氧菌.通过比较不同菌的相对数量变化发现五氯酚驯化后的颗粒污泥中产生了一系列对五氯酚降解有利的优势细菌和古菌,如Proteobacteria、Sphingomonas、Methanogenic bacterium等.     

微生物驱油物理模拟实验研究

根据物理模拟实验的相似性准则,进行了微生物在多孔介质中运移实验、微生物驱油提高采收率实验、不同的代谢产物对采收率的影响实验及复配菌种的增殖作用对采收率的影响实验,结果表明,N80菌种能够随着注入水在多孔介质中生长和运移,其代谢产物各组分对采收率影响随代谢产物浓度的增加而增加,对采收率提高值贡献最大的是代谢产物中的表面活性剂,其次为代谢产生的酸及气体,并发现单个代谢产物的驱油效率的代数和小于各组分共同作用时的驱油效率,表明微生物代谢产物中各组分间的协同作用有助于提高采收率。