Structural determinants of stereospecificity in yeast alcohol dehydrogenase.

Abstract:

:Replacing Leu-182 by Ala in yeast alcohol dehydrogenase (YADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1) yields a mutant that retains 34% of its kcat value and makes one stereochemical "mistake" every 850,000 turnovers (instead of approximately 1 error every 7,000,000,000 turnovers in native YADH) in its selection of the 4-Re hydrogen of NADH. Half of the decrease in stereochemical fidelity comes from an increase in the rate of transfer of the 4-Si hydrogen of NADH. The mutant also accepts 5-methylnicotinamide adenine dinucleotide, a cofactor analog not accepted by native YADH. The stereospecificity of the mutant is lower still with analogs of NADH where the carboxamide group of the nicotinamide ring is replaced by groups with weaker hydrogen bonding potential. For example, with thio-NADH, the mutant enzyme makes 1 stereochemical "mistake" every 450 turnovers. Finally, the double mutant T157S/L182A, in which Thr-157 is replaced by Ser and Leu-182 is replaced by Ala, also shows decreased stereochemical fidelity. These results suggest that Si transfer in the mutant enzymes arises from NADH bound in a syn conformation in the active site and that this binding is not obstructed in native YADH by side chains essential for catalysis.

authors

Weinhold EG,Glasfeld A,Ellington AD,Benner SA

doi

10.1073/pnas.88.19.8420

subject

Has Abstract

pub_date

1991-10-01 00:00:00

pages

8420-4

issue

19

eissn

0027-8424

issn

1091-6490

journal_volume

88

pub_type

杂志文章
  • Mechanism of integrating foreign DNA during transformation of Bacillus subtilis.

    abstract::Genes encoding thymidylate synthetase from Bacillus subtilis bacteriophages were cloned in Escherichia coli. Chimeric plasmids pCD1 and pCD3 were constructed from site-specific endonuclease digests of bacteriophage phi3T DNA cloned in pMB9 in E. coli. Similar cloning techniques with bacteriophage beta22 DNA yielded ch...

    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.75.8.3664

    authors: Duncan CH,Wilson GA,Young FE

    更新日期:1978-08-01 00:00:00

  • Remodeling of synaptic architecture during hippocampal "kindling".

    abstract::The "kindling" phenomenon is associated with long-lasting facilitation of synaptic transmission. A possible mechanism of such facilitation could involve changes in the number of synaptic contacts. However, previous attempts to demonstrate a synaptic morphological alteration that could account for the long-term effects...

    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.85.9.3260

    authors: Geinisman Y,Morrell F,deToledo-Morrell L

    更新日期:1988-05-01 00:00:00

  • Partial D-amino acid substitution: Improved enzymatic stability and preserved Ab recognition of a MUC2 epitope peptide.

    abstract::The stability of an immunogen against enzymatic degradation is considered an important factor for the design of synthetic vaccines. For our studies, we have selected an epitope from the tandem-repeat unit of the high-molecular-weight MUC2 mucin glycoprotein, which can be underglycosylated in case of colon cancer. In 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.0407677102

    authors: Tugyi R,Uray K,Iván D,Fellinger E,Perkins A,Hudecz F

    更新日期:2005-01-11 00:00:00

  • Trophic and size structure of West Indian bird communities.

    abstract::Much discussion has occurred in recent years on whether observed patterns of structure in island bird communities are the result of competitive interactions among species or independent rates of colonization and extinction. Here two patterns of structure are presented for birds on 12 West Indian islands. Each of four ...

    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.79.5.1563

    authors: Faaborg J

    更新日期:1982-03-01 00:00:00

  • Mechanism of glucoregulatory responses to stress and their deficiency in diabetes.

    abstract::During exercise, increased energy demands are met by increased glucose production that occurs simultaneously with the increased glucose uptake. We had previously observed that, during exercise, metabolic clearance rate of glucose (MCR) increases markedly in normal, but only marginally in poorly controlled diabetic dog...

    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.88.4.1296

    authors: Miles PD,Yamatani K,Lickley HL,Vranic M

    更新日期:1991-02-15 00:00:00

  • TALEN-engineered AR gene rearrangements reveal endocrine uncoupling of androgen receptor in prostate cancer.

    abstract::Androgen receptor (AR) target genes direct development and survival of the prostate epithelial lineage, including prostate cancer (PCa). Thus, endocrine therapies that inhibit the AR ligand-binding domain (LBD) are effective in treating PCa. AR transcriptional reactivation is central to resistance, as evidenced by the...

    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.1308587110

    authors: Nyquist MD,Li Y,Hwang TH,Manlove LS,Vessella RL,Silverstein KA,Voytas DF,Dehm SM

    更新日期:2013-10-22 00:00:00

  • AMPA receptors are exocytosed in stimulated spines and adjacent dendrites in a Ras-ERK-dependent manner during long-term potentiation.

    abstract::The exocytosis of AMPA receptors is a key step in long-term potentiation (LTP), yet the timing and location of exocytosis and the signaling pathways involved in exocytosis during synaptic plasticity are not fully understood. Here we combine two-photon uncaging with two-photon imaging of a fluorescent label of surface ...

    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.0913875107

    authors: Patterson MA,Szatmari EM,Yasuda R

    更新日期:2010-09-07 00:00:00

  • Toward controlling gene expression at will: specific regulation of the erbB-2/HER-2 promoter by using polydactyl zinc finger proteins constructed from modular building blocks.

    abstract::To create a universal system for the control of gene expression, we have studied methods for the construction of novel polydactyl zinc finger proteins that recognize extended DNA sequences. Elsewhere we have described the generation of zinc finger domains recognizing sequences of the 5'-GNN-3' subset of a 64-member zi...

    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.95.25.14628

    authors: Beerli RR,Segal DJ,Dreier B,Barbas CF 3rd

    更新日期:1998-12-08 00:00:00

  • Influence of chromosomal integration on glucocorticoid-regulated transcription of growth-stimulating papillomavirus genes E6 and E7 in cervical carcinoma cells.

    abstract::In most cervical carcinoma cells the E6 and E7 genes of specific human papillomaviruses are transcribed from viral sequences integrated into host cell chromosomes. Glucocorticoids activate the promoter elements of various human papillomaviruses in transient-expression assays. We have analyzed the effect of dexamethaso...

    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.88.4.1411

    authors: von Knebel Doeberitz M,Bauknecht T,Bartsch D,zur Hausen H

    更新日期:1991-02-15 00:00:00

  • Human biliverdin reductase is an ERK activator; hBVR is an ERK nuclear transporter and is required for MAPK signaling.

    abstract::Activation of the MEK/ERK/Elk-signaling cascade is a mechanism for relaying mitogenic and stress stimuli for gene activation. MEK1 is the proximate kinase for activation of ERK1/2, and nuclear targeting of ERK1/2 is obligatory for Elk1 transcriptional activity. Human biliverdin reductase (hBVR) is a recently described...

    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.0800750105

    authors: Lerner-Marmarosh N,Miralem T,Gibbs PE,Maines MD

    更新日期:2008-05-13 00:00:00

  • Components of histidine transport: histidine-binding proteins and hisP protein.

    abstract::The high-affinity (K(m) = 3 x 10(-8) M) transport system for histidine in Salmonella typhimurium has been resolved into three components: J, K, and P. J, which is a histidine-binding protein released by osmotic shock, is specified by the hisJ gene: hisJ mutants lack the binding protein and are defective in histidine 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.66.4.1096

    authors: Ames GF,Lever J

    更新日期:1970-08-01 00:00:00

  • Fossil vertebrates from Antigua, Lesser Antilles: Evidence for late Holocene human-caused extinctions in the West Indies.

    abstract::Vertebrate remains recovered from a limestone fissure filling on Antigua, Lesser Antilles, are associated with radiocarbon dates ranging from 4300 to 2500 yr B.P., contemporaneous with the earliest aboriginal human occupation of the island. Nine taxa of lizards, snakes, birds, bats, and rodents (one-third of the total...

    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.81.14.4448

    authors: Steadman DW,Pregill GK,Olson SL

    更新日期:1984-07-01 00:00:00

  • Membrane texture induced by specific protein binding and receptor clustering: active roles for lipids in cellular function.

    abstract::Biological membranes are complex, self-organized structures that define boundaries and compartmentalize space in living matter. Composed of a wide variety of lipid and protein molecules, these responsive surfaces mediate transmembrane signaling and material transport within the cell and with its environment. It is wel...

    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.1014579108

    authors: Watkins EB,Miller CE,Majewski J,Kuhl TL

    更新日期:2011-04-26 00:00:00

  • The antiinflammatory effect of laminar flow: the role of PPARgamma, epoxyeicosatrienoic acids, and soluble epoxide hydrolase.

    abstract::We previously reported that laminar flow activates peroxisome proliferator-activated receptor gamma (PPARgamma) in vascular endothelial cells in a ligand-dependent manner that involves phospholipase A2 and cytochrome P450 epoxygenases. In this study, we investigated whether epoxyeicosatrienoic acids (EETs), the cataly...

    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.0508081102

    authors: Liu Y,Zhang Y,Schmelzer K,Lee TS,Fang X,Zhu Y,Spector AA,Gill S,Morisseau C,Hammock BD,Shyy JY

    更新日期:2005-11-15 00:00:00

  • Targeted gene disruption of methionine aminopeptidase 2 results in an embryonic gastrulation defect and endothelial cell growth arrest.

    abstract::The antiangiogenic agent fumagillin (Fg) and its analog TNP-470 bind to intracellular metalloprotease methionine aminopeptidase-2 (MetAP-2) and inhibit endothelial cell growth in a p53-dependent manner. To confirm the role of MetAP-2 in endothelial cell proliferation and to validate it as a physiological target for th...

    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.0511313103

    authors: Yeh JJ,Ju R,Brdlik CM,Zhang W,Zhang Y,Matyskiela ME,Shotwell JD,Crews CM

    更新日期:2006-07-05 00:00:00

  • Dynamics of nitric oxide controlled by protein complex in bacterial system.

    abstract::Nitric oxide (NO) plays diverse and significant roles in biological processes despite its cytotoxicity, raising the question of how biological systems control the action of NO to minimize its cytotoxicity in cells. As a great example of such a system, we found a possibility that NO-generating nitrite reductase (NiR) f...

    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.1621301114

    authors: Terasaka E,Yamada K,Wang PH,Hosokawa K,Yamagiwa R,Matsumoto K,Ishii S,Mori T,Yagi K,Sawai H,Arai H,Sugimoto H,Sugita Y,Shiro Y,Tosha T

    更新日期:2017-09-12 00:00:00

  • Structural insights into the N-terminal GIY-YIG endonuclease activity of Arabidopsis glutaredoxin AtGRXS16 in chloroplasts.

    abstract::Glutaredoxins (Grxs) have been identified across taxa as important mediators in various physiological functions. A chloroplastic monothiol glutaredoxin, AtGRXS16 from Arabidopsis thaliana, comprises two distinct functional domains, an N-terminal domain (NTD) with GlyIleTyr-TyrIleGly (GIY-YIG) endonuclease motif and a ...

    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.1306899110

    authors: Liu X,Liu S,Feng Y,Liu JZ,Chen Y,Pham K,Deng H,Hirschi KD,Wang X,Cheng N

    更新日期:2013-06-04 00:00:00

  • Specific disruption of vimentin filament organization in monkey kidney CV-1 cells by diphtheria toxin, exotoxin A, and cycloheximide.

    abstract::We have examined the effect of diphtheria toxin, Pseudomonas aeruginosa exotoxin A, and cycloheximide on the CV-1 cell cytoskeleton. Within a few hours after producing an inhibition of cellular protein synthesis, all these agents specifically disrupted the organization of the vimentin filament system with no discernab...

    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.77.12.7267

    authors: Sharpe AH,Chen LB,Murphy JR,Fields BN

    更新日期:1980-12-01 00:00:00

  • In vitro transcription of heat-shock-specific RNA from chromatin of Drosophila melanogaster cells.

    abstract::Polyadenylylated RNA synthesized after heat shock was isolated from polysomes of cultured cells of Drosophila melanogaster and used as template to prepare cDNA. An excess of poly(A)-RNA from heat-shocked cells hybridized to 80% of the cDNA, whereas cytoplasmic poly(A)-RNA from cells grown at 25 degrees could drive onl...

    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.75.2.759

    authors: Biessmann H,Levy B,McCarthy BJ

    更新日期:1978-02-01 00:00:00

  • Navigating transformations in governance of Chilean marine coastal resources.

    abstract::Marine ecosystems are in decline. New transformational changes in governance are urgently required to cope with overfishing, pollution, global changes, and other drivers of degradation. Here we explore social, political, and ecological aspects of a transformation in governance of Chile's coastal marine resources, from...

    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.1012021107

    authors: Gelcich S,Hughes TP,Olsson P,Folke C,Defeo O,Fernández M,Foale S,Gunderson LH,Rodríguez-Sickert C,Scheffer M,Steneck RS,Castilla JC

    更新日期:2010-09-28 00:00:00

  • Localized maternal mRNA related to transforming growth factor beta mRNA is concentrated in a cytokeratin-enriched fraction from Xenopus oocytes.

    abstract::The localized maternal RNA Vg1 resides in the cortical region of the vegetal pole of fully grown Xenopus oocytes and is inherited by only a subset of blastomeres in the early embryo [Weeks, D. L. & Melton, D. A. (1987) Cell 51, 861-867]. Because RNA-cytoskeletal interactions may play a role in RNA localization, 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.85.20.7612

    authors: Pondel MD,King ML

    更新日期:1988-10-01 00:00:00

  • Synthesis, bacterial expression, and mutagenesis of the gene coding for mammalian cytochrome b5.

    abstract::We have totally synthesized a gene that codes for rat hepatic cytochrome b5. The 5' flanking region was designed for efficient expression of this gene in Escherichia coli by incorporating an optimum ribosome binding site and spacer region. Both a soluble form, analogous to the protease-treated microsomal protein, as w...

    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.83.24.9443

    authors: Beck von Bodman S,Schuler MA,Jollie DR,Sligar SG

    更新日期:1986-12-01 00:00:00

  • Integration of epigenetic and genetic controls of seed size by cytokinin in Arabidopsis.

    abstract::The development of seeds in flowering plants is placed under complex interactions between maternal tissues, the embryo, and the endosperm. The endosperm plays a major role in the regulation of seed size. In Arabidopsis thaliana, endosperm size depends on the coordination of the genetic pathway HAIKU (IKU) with epigene...

    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.1305175110

    authors: Li J,Nie X,Tan JL,Berger F

    更新日期:2013-09-17 00:00:00

  • Predicting protein function from structure: unique structural features of proteases.

    abstract::We have noted consistent structural similarities among unrelated proteases. In comparison with other proteins of similar size, proteases have smaller than average surface areas, smaller radii of gyration, and higher C(alpha) densities. These findings imply that proteases are, as a group, more tightly packed than other...

    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.070548997

    authors: Stawiski EW,Baucom AE,Lohr SC,Gregoret LM

    更新日期:2000-04-11 00:00:00

  • Old World sources of the first New World human inhabitants: a comparative craniofacial view.

    abstract::Human craniofacial data were used to assess the similarities and differences between recent and prehistoric Old World samples, and between these samples and a similar representation of samples from the New World. The data were analyzed by the neighbor-joining clustering procedure, assisted by bootstrapping and by cano...

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

    pub_type: 历史文章,杂志文章

    doi:10.1073/pnas.171305898

    authors: Brace CL,Nelson AR,Seguchi N,Oe H,Sering L,Qifeng P,Yongyi L,Tumen D

    更新日期:2001-08-14 00:00:00

  • Loss of p63 and its microRNA-205 target results in enhanced cell migration and metastasis in prostate cancer.

    abstract::p63 inhibits metastasis. Here, we show that p63 (both TAp63 and ΔNp63 isoforms) regulates expression of miR-205 in prostate cancer (PCa) cells, and miR-205 is essential for the inhibitory effects of p63 on markers of epithelial-mesenchymal transition (EMT), such as ZEB1 and vimentin. Correspondingly, the inhibitory ef...

    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.1110977109

    authors: Tucci P,Agostini M,Grespi F,Markert EK,Terrinoni A,Vousden KH,Muller PA,Dötsch V,Kehrloesser S,Sayan BS,Giaccone G,Lowe SW,Takahashi N,Vandenabeele P,Knight RA,Levine AJ,Melino G

    更新日期:2012-09-18 00:00:00

  • Climatic change and wetland desiccation cause amphibian decline in Yellowstone National Park.

    abstract::Amphibians are a bellwether for environmental degradation, even in natural ecosystems such as Yellowstone National Park in the western United States, where species have been actively protected longer than anywhere else on Earth. We document that recent climatic warming and resultant wetland desiccation are causing sev...

    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.0809090105

    authors: McMenamin SK,Hadly EA,Wright CK

    更新日期:2008-11-04 00:00:00

  • Heat transfer from nanoparticles: a corresponding state analysis.

    abstract::In this contribution, we study situations in which nanoparticles in a fluid are strongly heated, generating high heat fluxes. This situation is relevant to experiments in which a fluid is locally heated by using selective absorption of radiation by solid particles. We first study this situation for different types 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.0901372106

    authors: Merabia S,Shenogin S,Joly L,Keblinski P,Barrat JL

    更新日期:2009-09-08 00:00:00

  • Late Jurassic salamandroid from western Liaoning, China.

    abstract::A Jurassic salamander, Beiyanerpeton jianpingensis (gen. et sp. nov.), from a recently found site in western Liaoning Province, China is the earliest known record of Salamandroidea. As a Late Jurassic record of the group, it extends the range of the clade by ~40 Ma. The Late Jurassic taxon is neotenic and represented ...

    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.1009828109

    authors: Gao KQ,Shubin NH

    更新日期:2012-04-10 00:00:00

  • The sigma factor sigma s affects antibiotic production and biological control activity of Pseudomonas fluorescens Pf-5.

    abstract::Pseudomonas fluorescens Pf-5, a rhizosphere-inhabiting bacterium that suppresses several soilborne pathogens of plants, produces the antibiotics pyrrolnitrin, pyoluteorin, and 2,4-diacetylphloroglucinol. A gene necessary for pyrrolnitrin production by Pf-5 was identified as rpoS, which encodes the stationary-phase sig...

    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.92.26.12255

    authors: Sarniguet A,Kraus J,Henkels MD,Muehlchen AM,Loper JE

    更新日期:1995-12-19 00:00:00