共查询到20条相似文献，搜索用时 511 毫秒
1.
The homodimeric mammalian fatty acid synthase is one of the most complex cellular multienzymes, in that each 270-kilodalton polypeptide chain carries all seven functional domains required for fatty acid synthesis. We have calculated a 4.5 angstrom-resolution x-ray crystallographic map of porcine fatty acid synthase, highly homologous to the human multienzyme, and placed homologous template structures of all individual catalytic domains responsible for the cyclic elongation of fatty acid chains into the electron density. The positioning of domains reveals the complex architecture of the multienzyme forming an intertwined dimer with two lateral semicircular reaction chambers, each containing a full set of catalytic domains required for fatty acid elongation. Large distances between active sites and conformational differences between the reaction chambers demonstrate that mobility of the acyl carrier protein and general flexibility of the multienzyme must accompany handover of the reaction intermediates during the reaction cycle. 相似文献
2.
Jenni S Leibundgut M Boehringer D Frick C Mikolásek B Ban N 《Science (New York, N.Y.)》2007,316(5822):254-261
We report crystal structures of the 2.6-megadalton alpha6beta6 heterododecameric fatty acid synthase from Thermomyces lanuginosus at 3.1 angstrom resolution. The alpha and beta polypeptide chains form the six catalytic domains required for fatty acid synthesis and numerous expansion segments responsible for extensive intersubunit connections. Detailed views of all active sites provide insights into substrate specificities and catalytic mechanisms and reveal their unique characteristics, which are due to the integration into the multienzyme. The mode of acyl carrier protein attachment in the reaction chamber, together with the spatial distribution of active sites, suggests that iterative substrate shuttling is achieved by a relatively restricted circular motion of the carrier domain in the multifunctional enzyme. 相似文献
3.
Mammalian fatty acid synthase is a large multienzyme that catalyzes all steps of fatty acid synthesis. We have determined its crystal structure at 3.2 angstrom resolution covering five catalytic domains, whereas the flexibly tethered terminal acyl carrier protein and thioesterase domains remain unresolved. The structure reveals a complex architecture of alternating linkers and enzymatic domains. Substrate shuttling is facilitated by flexible tethering of the acyl carrier protein domain and by the limited contact between the condensing and modifying portions of the multienzyme, which are mainly connected by linkers rather than direct interaction. The structure identifies two additional nonenzymatic domains: (i) a pseudo-ketoreductase and (ii) a peripheral pseudo-methyltransferase that is probably a remnant of an ancestral methyltransferase domain maintained in some related polyketide synthases. The structural comparison of mammalian fatty acid synthase with modular polyketide synthases shows how their segmental construction allows the variation of domain composition to achieve diverse product synthesis. 相似文献
4.
Mechanism of the selective catalytic reduction of nitric oxide by ammonia elucidated by in situ on-line fourier transform infrared spectroscopy 总被引：1，自引：0，他引：1
Topsøe NY 《Science (New York, N.Y.)》1994,265(5176):1217-1219
The selective catalytic reduction reaction of nitric oxide bv ammonia over vanadia-titania catalysts is one of the methods of removing NOx pollution. In the present study, it has been possible to identify the reaction mechanism and the nature of the active sites in these catalysts by combining transient or steady-state in situ (Fourier transform infrared spectroscopy) experiments directly with on-line activity studies. The results suggest a catalytic cycle that consists of both acid and redox reactions and involves both surface V-OH (Br?nsted acid sites) and V=O species. A fundamental microkinetic model is proposed, which accounts for the observed industrial kinetics performance. 相似文献
5.
[目的]研究粗蓖麻油(含水、游离脂肪酸)与甲醇为原料的生物柴油合成体系的反应热力学特性。[方法]采用Yoneda基团贡献法和其他经验公式计算了组成蓖麻油的5种脂肪酸甘油三酸酯(TG)、甘油二酸酯(DG)、甘油单酸酯(MG)、脂肪酸甲酯(ME)及游离脂肪酸(FFA)的焓、熵及热容等基础热力学数据;对生物柴油合成体系,反应涉及酯交换、酯化及水解共3类7个反应进行了热力学分析。[结果]蓖麻油与甲醇的第1步和第3步酯交换反应的吉布斯自由能变化(△rG0)均小于0,第二步酯交换反应的△rG0大于0,游离脂肪酸与甲醇的酯化反应的△rG0小于0,蓖麻油与水的3步水解反应的△rG0均大于0。[结论]蓖麻油中游离脂肪酸的存在比水的存在对蓖麻油的酯交换反应影响更大。 相似文献
6.
In the multifunctional fungal fatty acid synthase (FAS), the acyl carrier protein (ACP) domain shuttles reaction intermediates covalently attached to its prosthetic phosphopantetheine group between the different enzymatic centers of the reaction cycle. Here, we report the structure of the Saccharomyces cerevisiae FAS determined at 3.1 angstrom resolution with its ACP stalled at the active site of ketoacyl synthase. The ACP contacts the base of the reaction chamber through conserved, charge-complementary surfaces, which optimally position the ACP toward the catalytic cleft of ketoacyl synthase. The conformation of the prosthetic group suggests a switchblade mechanism for acyl chain delivery to the active site of the enzyme. 相似文献
7.
【目的】亚油酸是一种人体必需的脂肪酸,能加快体内脂肪代谢与分解、减少胆固醇在血管壁上形成积存、有效预防动脉硬化的发生、提高人体免疫力、促进骨骼发育、提高记忆力、预防脑部功能退化等功能。利用基因编辑技术对控制脂肪酸合成途径的关键酶ω-3脂肪酸去饱和酶基因进行精准编辑,关闭下游产物的合成途径,以便提高水稻上游亚油酸的含量,为创制富集亚油酸水稻材料提供依据。【方法】水稻脂肪酸合成途径中的关键酶ω-3脂肪酸去饱和酶基因（OsFAD3）存在2个拷贝,分别分布在第11和第12染色体,且两者cDNA同源性达97.32%。根据基因编辑原理（编辑的特异性取决于引导RNA（gRNA）的特性）,在2个同源基因序列高度一致的外显子区域,分别设计合成2个gRNA,并分别构建植物基因编辑载体,利用农杆菌介导法转化本地受体材料（富源4号）。通过对基因编辑植株进行靶位点测序鉴定,分析2个位点的编辑效率和基因型。同时,对OsFAD3双突变体进行粒宽、粒长等主要籽粒农艺性状分析。采用气质联用检测法测量OsFAD3双突变体籽粒的37种脂肪酸含量。【结果】获得2个位点的纯合编辑材料,同时也获得了其他不同基因型组合的编辑材料;与对照材料相比,OsFAD3双突变体的粒宽、粒长等主要籽粒农艺性状均没有发生显著变化,但稻谷中亚油酸的相对含量提高了3.36个百分点。【结论】在不改变籽粒主要农艺性状的前提下,实现了利用基因编辑技术通过转化一个载体同时精准敲除2个ω-3脂肪酸去饱和酶基因,提高了籽粒中亚油酸的相对含量。 相似文献
8.
【目的】研究不同营养水平对湖羊黄体期血液理化指标及卵泡发育的影响。【方法】选择28只经产母羊,于发情周期第6天分别按0.5倍体重维持需要量（R组）,1倍体重维持需要量（C组）和1.5倍体重维持需要量（S组）饲喂6 d,第12天每组屠宰6头;剩下的湖羊用于观察发情;分别于发情周期第7、8、10和12天采血。【结果】随着营养水平的提高,≥3.5 mm的卵泡数量显著增加（P＜0.05）、2.5-3.5 mm的卵泡数量显著降低（P＜0.05）,平均发情周期缩短（P＜0.05）,血液尿素、胆固醇和游离脂肪酸含量显著下降（P＜0.05）,甘油三酯含量显著提高（P＜0.05）;S组乳酸脱氢酶活性显著高于C组（P＜0.05）;血液尿酸、血氨、高密度脂蛋白、低密度脂蛋白浓度和谷草转氨酶和谷丙转氨酶活性差异不显著（P＞0.05）,但尿酸、血氨、低密度脂蛋白含量和谷草转氨酶活性具有时间效应（P＜0.05）。【结论】绵羊黄体期不同生理阶段具有不同的营养需求和代谢特点,黄体期限饲抑制卵泡发育与蛋白质和脂类合成降低、分解增强有关。 相似文献
9.
Behrens M Studt F Kasatkin I Kühl S Hävecker M Abild-Pedersen F Zander S Girgsdies F Kurr P Kniep BL Tovar M Fischer RW Nørskov JK Schlögl R 《Science (New York, N.Y.)》2012,336(6083):893-897
One of the main stumbling blocks in developing rational design strategies for heterogeneous catalysis is that the complexity of the catalysts impairs efforts to characterize their active sites. We show how to identify the crucial atomic structure motif for the industrial Cu/ZnO/Al(2)O(3) methanol synthesis catalyst by using a combination of experimental evidence from bulk, surface-sensitive, and imaging methods collected on real high-performance catalytic systems in combination with density functional theory calculations. The active site consists of Cu steps decorated with Zn atoms, all stabilized by a series of well-defined bulk defects and surface species that need to be present jointly for the system to work. 相似文献
10.
【目的】分析黄瓜基因组中与叶酸合成代谢相关的基因数量、定位以及表达特征,对关键酶基因进行生物信息学分析与克隆,旨在为黄瓜叶酸合成调控研究奠定基础。【方法】根据已报道的拟南芥叶酸合成相关基因,利用黄瓜基因组数据库中9930_V3版本进行BLAST比对。利用MapChart绘制黄瓜染色体物理图谱并对基因定位。利用qRT-PCR分析这些基因在黄瓜果实发育不同时期和不同材料中的表达量。通过MEGA、WebLOGO、ExPASy等工具对关键酶基因进行生物信息学分析。通过PCR扩增对关键酶基因进行克隆,并测序分析基因的序列差异。【结果】同源比对获得19个黄瓜叶酸代谢相关基因,这些基因不均匀分布在黄瓜7条染色体上,且以Chr.4和Chr.5上分布最多。通过对其中11个调控叶酸合成的基因在测序黄瓜9930果实发育不同时期以及果实叶酸含量高低差异显著的2份材料的表达量分析,发现CsFPGS、CsHPPK/CsDHPS和CsDHNA 3个基因与果实叶酸含量变化趋势完全一致;CsADCS、CsADCL、CsDHNA、CsHPPK/CsDHFS、CsFPGS、CsDHFS等基因的表达量在2份材料中具有显著差异。通过对2个调控叶酸合成限速步骤的关键酶基因CsGCHI和CsADCS的蛋白序列及蛋白结构域分析,发现各物种中CsGCHI的同源基因均具有2个GTP_cyclohydroI结构域;CsADCS的同源基因均具有2个GATase结构域、1个Anth_synt_I_N结构域和1个Chorismate_bind结构域。它们在不同物种中高度保守,进化树分析亲缘关系近的物种聚类到一起。分别扩增黄瓜果实低叶酸含量自交系65G和高叶酸含量自交系02245中CsGCHI和CsADCS的同源基因,序列分析表明CsaV3_1G041250全长为3 012 bp,CDS序列长度为1 413 bp,3个SNP位点的突变导致了氨基酸序列的变异;CsaV3_7G026240全长为3 047 bp,CDS长度1 407 bp,序列无变异;CsaV3_5G036360全长7 941 bp,CDS序列长度为2 706 bp,序列无变异。【结论】鉴定出19个不均匀分布在7条染色体上的黄瓜叶酸代谢相关基因,基因CsFPGS、CsHPPK/CsDHPS 、CsDHNA和CsADCS是影响黄瓜果实叶酸含量变化、导致叶酸含量高低显著差异的关键基因,调控叶酸合成限速步骤的关键酶基因GCHI及ADCS功能相对保守,CsGCHI在65G、02245中有3个SNP位点的突变导致了氨基酸序列的差异。 相似文献
11.
Protein-DNA interactions in vivo upstream of a cell cycle-regulated human H4 histone gene 总被引：23，自引：0，他引：23
Cell cycle-dependent histone genes are transcribed at a basal level throughout the cell cycle, with a three- to fivefold increase during early S phase. Protein-DNA interactions in the 5' promoter region of a cell cycle-regulated human H4 histone gene have been analyzed at single-nucleotide resolution in vivo. This region contains two sites, with four potential protein-binding domains, at which the DNA is protected from reaction with dimethyl sulfate in cells and from digestion with deoxyribonuclease I in nuclei. These protein-DNA interactions persist during all phases of the cell cycle and dissociate with 0.16 to 0.2M sodium chloride. 相似文献
12.
本研究利用生物信息学方法,对从三七根抑制消减杂交cDNA文库中随机挑选的91个EST序列进行了分析。结果表明:91个克隆组装分析后有4个重叠群和83个单拷贝EST,代表了87个基因。已知功能的基因序列共58个,占66%,按基因功能分类为10类,其中大多基因与代谢途径和分泌途径有关。未知功能基因序列29个,占33%。对33个氨基酸序列可通读的EST进行跨膜结构和信号肽分析,结果表明:9个克隆有信号肽,其中3个克隆有跨膜结构,可能为膜蛋白,6个克隆可能为分泌蛋白。功能位点分析结果表明:大多数基因与胞外分泌,调节细胞凋亡和细胞周期,信号传导有关。功能结构域分析结果表明:除9个克隆没有预测到功能结构域外,其它克隆的结构域与信号传导,转录调控,电子传递,协迫,光合作用,蛋白折叠等有关。亚细胞定位大多数位于细胞质和细胞核。 相似文献
13.
Mandal M Lee M Barrick JE Weinberg Z Emilsson GM Ruzzo WL Breaker RR 《Science (New York, N.Y.)》2004,306(5694):275-279
We identified a previously unknown riboswitch class in bacteria that is selectively triggered by glycine. A representative of these glycine-sensing RNAs from Bacillus subtilis operates as a rare genetic on switch for the gcvT operon, which codes for proteins that form the glycine cleavage system. Most glycine riboswitches integrate two ligand-binding domains that function cooperatively to more closely approximate a two-state genetic switch. This advanced form of riboswitch may have evolved to ensure that excess glycine is efficiently used to provide carbon flux through the citric acid cycle and maintain adequate amounts of the amino acid for protein synthesis. Thus, riboswitches perform key regulatory roles and exhibit complex performance characteristics that previously had been observed only with protein factors. 相似文献
14.
6个鸡种腺苷单磷酸脱氨酶1基因克隆及序列分析 总被引：3，自引：0，他引：3
以肌苷酸合成代谢过程中主要的催化酶之一的鸡腺苷单磷酸脱氨酶1（adenosine monophosphate deaminase 1,AMPD1）基因作为候选基因,分析了其在6个鸡种中的序列多样性,发现在525 bp的片段中共存在10个多态位点,其中120位的A→G,355位的A→G的碱基变化仅在泰和乌骨鸡、北京油鸡、茶花鸡肌苷酸含量较高的鸡种中出现,推测这两个位点与肌苷酸含量密切相关。另外该基因与体重也可能具有相关性。 相似文献
15.
S Donadio M J Staver J B McAlpine S J Swanson L Katz 《Science (New York, N.Y.)》1991,252(5006):675-679
In Saccharopolyspora erythraea, the genes that govern synthesis of the polyketide portion of the macrolide antibiotic erythromycin are organized in six repeated units that encode fatty acid synthase (FAS)-like activities. Each repeated unit is designated a module, and two modules are contained in a single open reading frame. A model for the synthesis of this complex polyketide is proposed, where each module encodes a functional synthase unit and each synthase unit participates specifically in one of the six FAS-like elongation steps required for formation of the polyketide. In addition, genetic organization and biochemical order of events appear to be colinear. Evidence for the model is provided by construction of a selected mutant and by isolation of a polyketide of predicted structure. 相似文献
16.
贵妃鸡肌肉脂肪酸的GC-MS 分析 总被引：1，自引：0，他引：1
应用气相色谱-质谱（GC-MS）联合技术对贵妃鸡肌肉脂肪酸组分进行分析, 并计算各脂肪酸的相对百
分比含量结果表明贵妃鸡肌肉中共分离和鉴定出21 种脂肪酸其中饱和脂肪酸6 种,分别是豆蔻酸,十五烷酸,
棕榈酸,十七烷酸硬脂酸和花生酸,占总脂肪酸的28.61%,不饱和脂肪酸（UFA）15 种,分别是十四碳一烯酸,棕榈
油酸,油酸,亚油酸,十七碳烯酸9,12,15-十八碳三烯酸11-二十碳一烯酸11,14 二十碳二烯酸8,11,14-二十碳三
烯酸,花生四烯酸（AA）,芥酸二十二碳五烯酸（DPA）4,7,10,13,16,19-二十二碳六烯酸甲酯（DHA）,酌-亚麻酸,
11,14,17-二十碳三烯酸,占总脂肪酸含量的71.45%多不饱和脂肪酸（PUFAs）占总脂肪酸含量的25.08%,必需脂肪
酸占总脂肪酸含量的23.52% 相似文献
17.
Copper active sites play a major role in enzymatic activation of dioxygen. We trapped the copper-dioxygen complex in the enzyme peptidylglycine-alphahydroxylating monooxygenase (PHM) by freezing protein crystals that had been soaked with a slow substrate and ascorbate in the presence of oxygen. The x-ray crystal structure of this precatalytic complex, determined to 1.85-angstrom resolution, shows that oxygen binds to one of the coppers in the enzyme with an end-on geometry. Given this structure, it is likely that dioxygen is directly involved in the electron transfer and hydrogen abstraction steps of the PHM reaction. These insights may apply to other copper oxygen-activating enzymes, such as dopamine beta-monooxygenase, and to the design of biomimetic complexes. 相似文献
18.
猪源新城疫病毒SP13株的F基因克隆及序列分析 总被引：1，自引：0，他引：1
采用RT-PCR方法对猪源新城疫病毒(NDV)SP13株的F基因进行了扩增与克隆,并测定出F基因的核苷酸全序列,推导出氨基酸序列.F基因全长为1662 bp,单一的开放阅读框,编码553个氨基酸的长肽,裂解位点的氨基酸序列为112G-R-Q-G-R-L117,与弱毒株在这一区域的序列(112G-R/K-Q-G/S-R-L117)相符;F蛋白有6个潜在的糖基化位点和13个Cys残基位点,其疏水构型有3个强疏水区.通过同源率、系统发育、致病性、疏水性和抗原性等比较分析的结果表明,SP13与LaSota、Clone 30株不但同源性达到99.9%,而且在致病性、疏水性和抗原性等方面也极为相似. 相似文献
19.
20.
Crystal structure of Cd,Zn metallothionein 总被引：9，自引：0，他引：9
W F Furey A H Robbins L L Clancy D R Winge B C Wang C D Stout 《Science (New York, N.Y.)》1986,231(4739):704-710
The anomalous scattering data from five Cd in the native protein were used to determine the crystal structure of cadmium, zinc (Cd,Zn) metallothionein isoform II from rat liver. The structure of a 4-Cd cluster was solved by direct methods. A 2.3 A resolution electron density map was calculated by iterative single-wavelength anomalous scattering. The structure is folded into two domains. The amino terminal domain (beta) of residues 1 to 29 enfolds a three-metal cluster of one Cd and two Zn atoms coordinated by six terminal cysteine thiolate ligands and three bridging cysteine thiolates. The carboxyl terminal domain (alpha) of residues 30 to 61 enfolds a 4-Cd cluster coordinated by six terminal and five bridging cysteine thiolates. All seven metal sites have tetrahedral coordination geometry. The domains are roughly spherical, and the diameter is 15 to 20 A; there is limited contact between domains. The folding of alpha and beta is topologically similar but with opposite chirality. Redundant, short cysteine-containing sequences have similar roles in cluster formation in both alpha and beta. 相似文献