In vitro reconstitution and steady-state analysis of the fatty acid synthase from Escherichia coli.

Abstract:

:Microbial fatty acid derivatives are emerging as promising alternatives to fossil fuel derived transportation fuels. Among bacterial fatty acid synthases (FAS), the Escherichia coli FAS is perhaps the most well studied, but little is known about its steady-state kinetic behavior. Here we describe the reconstitution of E. coli FAS using purified protein components and report detailed kinetic analysis of this reconstituted system. When all ketosynthases are present at 1 μM, the maximum rate of free fatty acid synthesis of the FAS exceeded 100 μM/ min. The steady-state turnover frequency was not significantly inhibited at high concentrations of any substrate or cofactor. FAS activity was saturated with respect to most individual protein components when their concentrations exceeded 1 μM. The exceptions were FabI and FabZ, which increased FAS activity up to concentrations of 10 μM; FabH and FabF, which decreased FAS activity at concentrations higher than 1 μM; and holo-ACP and TesA, which gave maximum FAS activity at 30 μM concentrations. Analysis of the S36T mutant of the ACP revealed that the unusual dependence of FAS activity on holo-ACP concentration was due, at least in part, to the acyl-phosphopantetheine moiety. MALDI-TOF mass spectrometry analysis of the reaction mixture further revealed medium and long chain fatty acyl-ACP intermediates as predominant ACP species. We speculate that one or more of such intermediates are key allosteric regulators of FAS turnover. Our findings provide a new basis for assessing the scope and limitations of using E. coli as a biocatalyst for the production of diesel-like fuels.

authors

Yu X,Liu T,Zhu F,Khosla C

doi

10.1073/pnas.1110852108

subject

Has Abstract

pub_date

2011-11-15 00:00:00

pages

18643-8

issue

46

eissn

0027-8424

issn

1091-6490

pii

1110852108

journal_volume

108

pub_type

杂志文章
  • A large region (approximately equal to 95 kDa) of human factor VIII is dispensable for in vitro procoagulant activity.

    abstract::Factor VIII (antihemophilic factor) is a high molecular weight plasma glycoprotein that participates in the blood clotting cascade. The recent cloning and sequence analysis of the cDNA encoding human factor VIII revealed an obvious domain structure for the protein, which can be represented as A1-A2-B-A3-C1-C2. We now ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.83.16.5939

    authors: Toole JJ,Pittman DD,Orr EC,Murtha P,Wasley LC,Kaufman RJ

    更新日期:1986-08-01 00:00:00

  • A muscarone-binding material in electroplax and its relation to the acetylcholine receptor. I. Centrifugal assay.

    abstract::A muscarone-binding material has been found in Torpedo electroplax. The material behaves in several ways as one would expect of the binding component of the acetylcholine receptor, especially in its high affinity for muscarone (approximate K = 7 x 10(-6)M), acetylcholine (approximate K = 1.1 x 10(-6)M), curare, and ni...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.63.2.496

    authors: O'Brien RD,Gilmour LP

    更新日期:1969-06-01 00:00:00

  • Atoms of recognition in human and computer vision.

    abstract::Discovering the visual features and representations used by the brain to recognize objects is a central problem in the study of vision. Recently, neural network models of visual object recognition, including biological and deep network models, have shown remarkable progress and have begun to rival human performance in...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1513198113

    authors: Ullman S,Assif L,Fetaya E,Harari D

    更新日期:2016-03-08 00:00:00

  • H2O2-induced uncoupling of bovine lens Na+,K+-ATPase.

    abstract::A 1-hr exposure of bovine lenses in organ culture to H2O2 concentrations in the range found in the aqueous fluid of patients with cataracts inhibits 86Rb+ influx. At 1 mM H2O2, complete inhibition was observed and further investigated. Membrane permeability is slightly decreased. Although lactate concentrations increa...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.7.2044

    authors: Garner WH,Garner MH,Spector A

    更新日期:1983-04-01 00:00:00

  • Stepwise assembly of the Nova-regulated alternative splicing network in the vertebrate brain.

    abstract::Novel organismal structures in metazoans are often undergirded by complex gene regulatory networks; as such, understanding the emergence of new structures through evolution requires reconstructing the series of evolutionary steps leading to these underlying networks. Here, we reconstruct the step-by-step assembly of t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1012333108

    authors: Irimia M,Denuc A,Burguera D,Somorjai I,Martín-Durán JM,Genikhovich G,Jimenez-Delgado S,Technau U,Roy SW,Marfany G,Garcia-Fernàndez J

    更新日期:2011-03-29 00:00:00

  • Structure of factor-inhibiting hypoxia-inducible factor 1: An asparaginyl hydroxylase involved in the hypoxic response pathway.

    abstract::Precise regulation of the evolutionarily conserved hypoxia-inducible transcription factor (HIF) ensures proper adaptation to variations in oxygen availability throughout development and into adulthood. Oxygen-dependent regulation of HIF stability and activity are mediated by hydroxylation of conserved proline and aspa...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.202614999

    authors: Dann CE 3rd,Bruick RK,Deisenhofer J

    更新日期:2002-11-26 00:00:00

  • Gut-expressed gustducin and taste receptors regulate secretion of glucagon-like peptide-1.

    abstract::Glucagon-like peptide-1 (GLP-1), released from gut endocrine L cells in response to glucose, regulates appetite, insulin secretion, and gut motility. How glucose given orally, but not systemically, induces GLP-1 secretion is unknown. We show that human duodenal L cells express sweet taste receptors, the taste G protei...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0706890104

    authors: Jang HJ,Kokrashvili Z,Theodorakis MJ,Carlson OD,Kim BJ,Zhou J,Kim HH,Xu X,Chan SL,Juhaszova M,Bernier M,Mosinger B,Margolskee RF,Egan JM

    更新日期:2007-09-18 00:00:00

  • The maximal affinity of ligands.

    abstract::We explore the question of what are the best ligands for macromolecular targets. A survey of experimental data on a large number of the strongest-binding ligands indicates that the free energy of binding increases with the number of nonhydrogen atoms with an initial slope of approximately -1.5 kcal/mol (1 cal = 4.18 J...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.18.9997

    authors: Kuntz ID,Chen K,Sharp KA,Kollman PA

    更新日期:1999-08-31 00:00:00

  • Complex formation between stage-specific oocyte factors and a Xenopus mRNA localization element.

    abstract::It is a long-standing proposal that localization of maternal factors in eggs can provide the basis for pattern formation in the early embryo. The localized information can be stored as RNA, one example being Vg1 RNA, which is localized exclusively to the vegetal hemisphere of Xenopus oocytes and eggs. Localization of ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.25.14608

    authors: Mowry KL

    更新日期:1996-12-10 00:00:00

  • Intercellular trafficking of a KNOTTED1 green fluorescent protein fusion in the leaf and shoot meristem of Arabidopsis.

    abstract::Dominant mutations in the maize homeobox gene knotted1 (kn1) act nonautonomously during maize leaf development, indicating that Kn1 is involved in the generation or transmission of a developmental signal that passes from the inner layers of the leaf to epidermal cells. We previously found that this nonautonomous activ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.052484099

    authors: Kim JY,Yuan Z,Cilia M,Khalfan-Jagani Z,Jackson D

    更新日期:2002-03-19 00:00:00

  • The colorectal microRNAome.

    abstract::MicroRNAs (miRNAs) are a class of small noncoding RNAs that have important regulatory roles in multicellular organisms. The public miRNA database contains 321 human miRNA sequences, 234 of which have been experimentally verified. To explore the possibility that additional miRNAs are present in the human genome, we hav...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0511155103

    authors: Cummins JM,He Y,Leary RJ,Pagliarini R,Diaz LA Jr,Sjoblom T,Barad O,Bentwich Z,Szafranska AE,Labourier E,Raymond CK,Roberts BS,Juhl H,Kinzler KW,Vogelstein B,Velculescu VE

    更新日期:2006-03-07 00:00:00

  • Cell cycle-dependent initiation and lineage-dependent abrogation of GATA-1 expression in pure differentiating hematopoietic progenitors.

    abstract::The programmed activation/repression of transcription factors in early hematopoietic differentiation has not yet been explored. The DNA-binding protein GATA-1 is required for normal erythroid development and regulates erythroid-expressed genes in maturing erythroblasts. We analyzed GATA-1 expression in early human adu...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.89.14.6353

    authors: Sposi NM,Zon LI,Carè A,Valtieri M,Testa U,Gabbianelli M,Mariani G,Bottero L,Mather C,Orkin SH

    更新日期:1992-07-15 00:00:00

  • A universal protein-protein interaction motif in the eubacterial DNA replication and repair systems.

    abstract::The interaction between DNA polymerases and sliding clamp proteins confers processivity in DNA synthesis. This interaction is critical for most DNA replication machines from viruses and prokaryotes to higher eukaryotes. The clamp proteins also participate in a variety of dynamic and competing protein-protein interacti...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.191384398

    authors: Dalrymple BP,Kongsuwan K,Wijffels G,Dixon NE,Jennings PA

    更新日期:2001-09-25 00:00:00

  • PICK1 inhibits the E3 ubiquitin ligase activity of Parkin and reduces its neuronal protective effect.

    abstract::Parkin functions as a multipurpose E3 ubiquitin ligase, and Parkin loss of function is associated with both sporadic and familial Parkinson's disease (PD). We report that the Bin/Amphiphysin/Rvs (BAR) domain of protein interacting with PRKCA1 (PICK1) bound to the really interesting new gene 1 (RING1) domain of Parkin ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1716506115

    authors: He J,Xia M,Yeung PKK,Li J,Li Z,Chung KK,Chung SK,Xia J

    更新日期:2018-07-24 00:00:00

  • Effects of an HIV-1 maturation inhibitor on the structure and dynamics of CA-SP1 junction helices in virus-like particles.

    abstract::HIV-1 maturation involves conversion of the immature Gag polyprotein lattice, which lines the inner surface of the viral membrane, to the mature capsid protein (CA) lattice, which encloses the viral RNA. Maturation inhibitors such as bevirimat (BVM) bind within six-helix bundles, formed by a segment that spans the jun...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1917755117

    authors: Gupta S,Louis JM,Tycko R

    更新日期:2020-05-12 00:00:00

  • Inflammatory mediator-induced hypothalamic-pituitary-adrenal axis activation is defective in streptococcal cell wall arthritis-susceptible Lewis rats.

    abstract::Inbred Lewis (LEW/N) female rats develop an arthritis in response to group A streptococcal cell wall peptidoglycan polysaccharide (SCW), which mimics human rheumatoid arthritis. Histocompatible Fischer (F344/N) rats do not develop arthritis in response to the same SCW stimulus. To evaluate this difference in inflammat...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.7.2374

    authors: Sternberg EM,Hill JM,Chrousos GP,Kamilaris T,Listwak SJ,Gold PW,Wilder RL

    更新日期:1989-04-01 00:00:00

  • Transition from one revolving cluster to two revolving clusters in the ground-state rotational bands of nuclei in the lanthanon region.

    abstract::Whereas 234(92)U142 and other actinon nuclei have ground-state bands that indicate that each nucleus consists of a sphere and a single revolving cluster with constant composition and with only a steady increase in the moment of inertia with increase in J, the angular-momentum quantum number, many of the lanthanon grou...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.88.3.820

    authors: Pauling L

    更新日期:1991-02-01 00:00:00

  • Scd1 controls de novo beige fat biogenesis through succinate-dependent regulation of mitochondrial complex II.

    abstract::Preadipocytes can give rise to either white adipocytes or beige adipocytes. Owing to their distinct abilities in nutrient storage and energy expenditure, strategies that specifically promote "beiging" of adipocytes hold great promise for counterbalancing obesity and metabolic diseases. Yet, factors dictating the diffe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1914553117

    authors: Liu K,Lin L,Li Q,Xue Y,Zheng F,Wang G,Zheng C,Du L,Hu M,Huang Y,Shao C,Kong X,Melino G,Shi Y,Wang Y

    更新日期:2020-02-04 00:00:00

  • Chick cartilage collagen: a new type of alpha 1 chain not present in bone or skin of the species.

    abstract::Isolation and chromatography of collagen from chick cartilage revealed an excess of alpha1 relative to alpha2-chains. Characterization of the cyanogen bromide peptides from the alpha1 fraction indicated the presence of two homologous forms of alpha1. The results suggest that cartilage contains two collagens, one ident...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.64.4.1264

    authors: Miller EJ,Matukas VJ

    更新日期:1969-12-01 00:00:00

  • Complete nucleotide sequence of the gene for the specific glycoprotein (gp55) of Friend spleen focus-forming virus.

    abstract::The complete nucleotide sequence of the gene for the specific glycoprotein (gp55) of the polycythemic strain of Friend spleen focus-forming virus (SFFV) was derived from the cloned SFFV DNA intermediate. The gp55 gene is present within 1.4 kilobases of the 5' side of the 3'long terminal repeat sequence. The open readi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.13.3913

    authors: Amanuma H,Katori A,Obata M,Sagata N,Ikawa Y

    更新日期:1983-07-01 00:00:00

  • Optimal decision making and matching are tied through diminishing returns.

    abstract::How individuals make decisions has been a matter of long-standing debate among economists and researchers in the life sciences. In economics, subjects are viewed as optimal decision makers who maximize their overall reward income. This framework has been widely influential, but requires a complete knowledge of the rew...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1703440114

    authors: Kubanek J

    更新日期:2017-08-08 00:00:00

  • Real-time imaging of calcium influx in mammalian cerebellar Purkinje cells in vitro.

    abstract::Real-time visualization of intracellular calcium concentration ([Ca2+]i) changes in mammalian Purkinje cells in vitro, utilizing the dye Fura-2, indicates that calcium action potentials are generated in the dendritic tree and follow a particular activation sequence. During spontaneous oscillations or after direct curr...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.13.5084

    authors: Sugimori M,Llinás RR

    更新日期:1990-07-01 00:00:00

  • Muscle-regulated expression and determinants for neuromuscular junctional localization of the mouse RIalpha regulatory subunit of cAMP-dependent protein kinase.

    abstract::In skeletal muscle, transcription of the gene encoding the mouse type Ialpha (RIalpha) subunit of the cAMP-dependent protein kinase is initiated from the alternative noncoding first exons 1a and 1b. Here, we report that activity of the promoter upstream of exon 1a (Pa) depends on two adjacent E boxes (E1 and E2) in NI...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.081393598

    authors: Barradeau S,Imaizumi-Scherrer T,Weiss MC,Faust DM

    更新日期:2001-04-24 00:00:00

  • Object grouping based on real-world regularities facilitates perception by reducing competitive interactions in visual cortex.

    abstract::In virtually every real-life situation humans are confronted with complex and cluttered visual environments that contain a multitude of objects. Because of the limited capacity of the visual system, objects compete for neural representation and cognitive processing resources. Previous work has shown that such attentio...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章,随机对照试验

    doi:10.1073/pnas.1400559111

    authors: Kaiser D,Stein T,Peelen MV

    更新日期:2014-07-29 00:00:00

  • Neoplastic transformation and tumorigenesis by the human protooncogene MYC.

    abstract::Damage to the protooncogene MYC has been implicated in the genesis of diverse human tumors, but the tumorigenic potential of the isolated gene has been disputed. Here we report the use of a retroviral vector to test the potency of human MYC for neoplastic transformation in avian cells. We found that sustained and abun...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.87.6.2102

    authors: Ramsay GM,Moscovici G,Moscovici C,Bishop JM

    更新日期:1990-03-01 00:00:00

  • Adult and fetal human globin genes are expressed following chromosomal transfer into MEL cells.

    abstract::Somatic cell hybridization of mouse erythroleukemia (MEL) cells and HEL cells, a human erythroleukemia line that produces fetal (gamma) but fails to express adult (beta) globin, was used to test whether the expression of the two human globin genes is regulated cis or trans. An experimental approach using anti-human gl...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.82.3.780

    authors: Papayannopoulou TH,Lindsley D,Kurachi S,Lewison K,Hemenway T,Melis M,Anagnou NP,Najfeld V

    更新日期:1985-02-01 00:00:00

  • Interfacial self-assembly of a bacterial hydrophobin.

    abstract::The majority of bacteria in the natural environment live within the confines of a biofilm. The Gram-positive bacterium Bacillus subtilis forms biofilms that exhibit a characteristic wrinkled morphology and a highly hydrophobic surface. A critical component in generating these properties is the protein BslA, which form...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1419016112

    authors: Bromley KM,Morris RJ,Hobley L,Brandani G,Gillespie RM,McCluskey M,Zachariae U,Marenduzzo D,Stanley-Wall NR,MacPhee CE

    更新日期:2015-04-28 00:00:00

  • Macrofaunal control of microbial community structure in continental margin sediments.

    abstract::Through a process called "bioturbation," burrowing macrofauna have altered the seafloor habitat and modified global carbon cycling since the Cambrian. However, the impact of macrofauna on the community structure of microorganisms is poorly understood. Here, we show that microbial communities across bioturbated, but ge...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1917494117

    authors: Deng L,Bölsterli D,Kristensen E,Meile C,Su CC,Bernasconi SM,Seidenkrantz MS,Glombitza C,Lagostina L,Han X,Jørgensen BB,Røy H,Lever MA

    更新日期:2020-07-07 00:00:00

  • Selecting antagonistic antibodies that control differentiation through inducible expression in embryonic stem cells.

    abstract::Antibodies that modulate receptor function have great untapped potential in the control of stem cell differentiation. In contrast to many natural ligands, antibodies are stable, exquisitely specific, and are unaffected by the regulatory mechanisms that act on natural ligands. Here we describe an innovative system for ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1312062110

    authors: Melidoni AN,Dyson MR,Wormald S,McCafferty J

    更新日期:2013-10-29 00:00:00

  • Direct photoaffinity labeling of tubulin with colchicine.

    abstract::Ultraviolet irradiation of the [3H]colchicine-tubulin complex leads to direct photolabeling of tubulin with low but practicable efficiency. The bulk (70% to greater than 90%) of the labeling occurs on beta-tubulin and appears early after irradiation, whereas alpha-tubulin is labeled later. The labeling ratio of beta-t...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.88.7.2820

    authors: Wolff J,Knipling L,Cahnmann HJ,Palumbo G

    更新日期:1991-04-01 00:00:00