当前位置： SCI文献检索 > JOURNAL OF MOLECULAR BIOLOGY期刊下所有文献 > An improved system for the generation and analysis of mutant proteins containing unnatural amino acids in Saccharomyces cerevisiae.

An improved system for the generation and analysis of mutant proteins containing unnatural amino acids in Saccharomyces cerevisiae.

Abstract：

:We have previously described methodology that makes it possible to genetically encode a wide array of unnatural amino acids in both prokaryotic and eukaryotic organisms. Here, we report the systematic optimization of a Saccharomyces cerevisiae expression system for the production of mutant proteins containing unnatural amino acids. Modifications include significant increases in both the expression levels of the orthogonal Escherichia coli amber suppressor tRNA(CUA) and cognate aminoacyl-tRNA synthetase (aaRS) pair, and expression of the target protein gene using a strong transcriptional promoter, optimized codons and elevated plasmid copy numbers. With this new system, a number of unnatural amino acids, including the photocrosslinkers p-benzoylphenylalanine and p-azidophenylalanine, and the chemically reactive amino acids, p-acetylphenylalanine and p-propargyloxyphenylalanine, were incorporated into human superoxide dismutase (hSOD) in yeast in good yields (maximally approximately 6-8 mg/l of culture in most cases). Mass spectrometric analysis of the hSOD mutants was performed with high dynamic range using multiple reaction monitoring that provided new insights into the factors that control the fidelity of unnatural amino acid incorporation.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Chen S,Schultz PG,Brock A

doi

10.1016/j.jmb.2007.05.017

subject

Has Abstract

pub_date

2007-08-03 00:00:00

pages

112-22

issue

1

eissn

0022-2836

issn

1089-8638

pii

S0022-2836(07)00627-4

journal_volume

371

pub_type

杂志文章

相关文献

JOURNAL OF MOLECULAR BIOLOGY文献大全
  • Origin of ultraviolet damage in DNA.

    abstract：:A novel ultraviolet (u.v.) footprinting technique has been used to analyze the formation of u.v. photoproducts at 250 bases of a 5 S rRNA gene under conditions where the gene is either double or single-stranded. Because many more types of u.v. damage can be detected by the u.v. footprinting technique than has been pre...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/0022-2836(89)90120-4

    authors： Becker MM,Wang Z

    更新日期：1989-12-05 00:00:00

  • Structural and functional analysis of a human 7 S K RNA gene.

    abstract：:Using purified RNA from HeLa cells, we have synthesized and cloned a cDNA encoding an almost entire 7 S K RNA. This cDNA probe was used to isolate 7 S K RNA gene sequences from a human genomic library by high-stringency colony hybridization. In order to differentiate between functional genes and related sequences, we ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/0022-2836(87)90325-1

    authors： Krüger W,Benecke BJ

    更新日期：1987-05-05 00:00:00

  • Local and global cooperativity in the human alpha-lactalbumin molten globule.

    abstract：:NMR spectroscopy has been used to follow the urea-induced unfolding of the low pH molten globule states of a single-disulfide variant of human alpha-lactalbumin ([28-111] alpha-LA) and of two mutants, each with a single proline substitution in a helix. [28-111] alpha-LA forms a molten globule very similar to that form...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2004.02.045

    authors： Quezada CM,Schulman BA,Froggatt JJ,Dobson CM,Redfield C

    更新日期：2004-04-16 00:00:00

  • Structure-function defects of the TWINKLE linker region in progressive external ophthalmoplegia.

    abstract：:TWINKLE is the helicase at the mitochondrial DNA (mtDNA) replication fork in mammalian cells. Mutations in the PEO1 gene, which encodes TWINKLE, cause autosomal dominant progressive external ophthalmoplegia (AdPEO), a disorder associated with deletions in mtDNA. Here, we characterized seven different AdPEO-causing mut...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2008.01.035

    authors： Korhonen JA,Pande V,Holmlund T,Farge G,Pham XH,Nilsson L,Falkenberg M

    更新日期：2008-03-28 00:00:00

  • Crystal structure of a bZIP/DNA complex at 2.2 A: determinants of DNA specific recognition.

    abstract：:The X-ray structure of the GCN4-bZIP protein bound to DNA containing the ATF/CREB recognition sequence has been refined at 2.2 A. The water-mediated interactions between the basic domain and DNA are revealed, and combined with a more accurate description of the direct contacts, further clarify how binding specificity ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1995.0645

    authors： Keller W,König P,Richmond TJ

    更新日期：1995-12-08 00:00:00

  • Topological frustration and the folding of interleukin-1 beta.

    abstract：:The cytokine, interleukin-1beta (IL-1beta), adopts a beta-trefoil fold. It is known to be much slower folding than similarly sized proteins, despite having a low contact order. Proteins are sufficiently well designed that their folding is not dominated by local energetic traps. Therefore, protein models that encode on...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2005.11.074

    authors： Gosavi S,Chavez LL,Jennings PA,Onuchic JN

    更新日期：2006-03-31 00:00:00

  • The C-terminal region of cytoplasmic polyadenylation element binding protein is a ZZ domain with potential for protein-protein interactions.

    abstract：:Cytoplasmic polyadenylation element binding protein (CPEB) provides temporal and spatial control of protein synthesis required for early development and neuronal synaptic plasticity. CPEB regulates protein expression by inhibiting polyadenylation of selected mRNA transcripts, which prevents binding of the ribosome for...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2013.03.009

    authors： Merkel DJ,Wells SB,Hilburn BC,Elazzouzi F,Pérez-Alvarado GC,Lee BM

    更新日期：2013-06-12 00:00:00

  • Three-dimensional solid-state NMR study of a seven-helical integral membrane proton pump--structural insights.

    abstract：:Proteorhodopsin (PR) is a recently discovered ubiquitous eubacterial retinal-binding light-driven proton pump. Almost 1000 PR variants are widely distributed in species of marine and freshwater bacteria, suggesting PR's important photobiological role. PR is a typical seven-transmembrane alpha-helical membrane protein ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2009.01.011

    authors： Shi L,Ahmed MA,Zhang W,Whited G,Brown LS,Ladizhansky V

    更新日期：2009-03-06 00:00:00

  • Localization of L11 protein on the ribosome and elucidation of its involvement in EF-G-dependent translocation.

    abstract：:L11 protein is located at the base of the L7/L12 stalk of the 50 S subunit of the Escherichia coli ribosome. Because of the flexible nature of the region, recent X-ray crystallographic studies of the 50 S subunit failed to locate the N-terminal domain of the protein. We have determined the position of the complete L11...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.2001.4907

    authors： Agrawal RK,Linde J,Sengupta J,Nierhaus KH,Frank J

    更新日期：2001-08-24 00:00:00

  • Impaired folding of the mitochondrial small TIM chaperones induces clearance by the i-AAA protease.

    abstract：:The intermembrane space of mitochondria contains a dedicated chaperone network-the small translocase of the inner membrane (TIM) family-for the sorting of hydrophobic precursors. All small TIMs are defined by the presence of a twin CX(3)C motif and the monomeric proteins are stabilized by two intramolecular disulfide ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2012.09.019

    authors： Baker MJ,Mooga VP,Guiard B,Langer T,Ryan MT,Stojanovski D

    更新日期：2012-12-14 00:00:00

  • Different intrastrand and interstrand contributions to stacking account for roll variations at the alternating purine-pyrimidine sequences in A-DNA and A-RNA.

    abstract：:An explanation is suggested for the roll alternation between low and high values in A-type nucleic acid duplexes containing alternating sequences of purine and pyrimidine residues. The explanation combines two points. (1) Roll inevitably occurs in A-type duplexes due to geometrical reasons. (2) Intrastrand base stacki...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/0022-2836(91)80172-q

    authors： Sponer J,Kypr J

    更新日期：1991-10-05 00:00:00

  • Synthesis and activity of piperazine-containing antirhinoviral agents and crystal structure of SDZ 880-061 bound to human rhinovirus 14.

    abstract：:A series of antipicornaviral agents containing piperazinyl moieties was synthesized with the objective of obtaining a compound with a broad spectrum of antirhinovirus activity, high potency (< or = 0.003 microgram/ml), and low cytotoxicity (> or = 30 micrograms/ml). Five compounds of this series were evaluated in deta...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1996.0307

    authors： Oren DA,Zhang A,Nesvadba H,Rosenwirth B,Arnold E

    更新日期：1996-05-31 00:00:00

  • Phosphorylation and mutation of phospholamban alter physical interactions with the sarcoplasmic reticulum calcium pump.

    abstract：:Phospholamban physically interacts with the sarcoplasmic reticulum calcium pump (SERCA) and regulates contractility of the heart in response to adrenergic stimuli. We studied this interaction using electron microscopy of 2D crystals of SERCA in complex with phospholamban. In earlier studies, phospholamban oligomers we...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2010.11.014

    authors： Glaves JP,Trieber CA,Ceholski DK,Stokes DL,Young HS

    更新日期：2011-01-21 00:00:00

  • Flexibility and variability of TIMP binding: X-ray structure of the complex between collagenase-3/MMP-13 and TIMP-2.

    abstract：:The excessive activity of matrix metalloproteinases (MMPs) contributes to pathological processes such as arthritis, tumor growth and metastasis if not balanced by the tissue inhibitors of metalloproteinases (TIMPs). In arthritis, the destruction of fibrillar (type II) collagen is one of the hallmarks, with MMP-1 (coll...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2006.11.072

    authors： Maskos K,Lang R,Tschesche H,Bode W

    更新日期：2007-03-02 00:00:00

  • Translation initiation factor 3 regulates switching between different modes of ribosomal subunit joining.

    abstract：:Ribosomal subunit joining is a key checkpoint in the bacterial translation initiation pathway during which initiation factors (IFs) regulate association of the 30S initiation complex (IC) with the 50S subunit to control formation of a 70S IC that can enter into the elongation stage of protein synthesis. The GTP-bound ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2014.09.024

    authors： MacDougall DD,Gonzalez RL Jr

    更新日期：2015-05-08 00:00:00

  • The critical roles of residues P235 and F236 of subunit A of the motor protein A-ATP synthase in P-loop formation and nucleotide binding.

    abstract：:The mutants P235A and F236A have been generated and their crystal structure was determined to resolutions of 2.38 and 2.35 A, respectively, in order to understand the residues involved in the formation of the novel arched P-loop of subunit A of the A-ATP synthase from Pyrococcus horikoshii OT3. Both the structures sho...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2010.06.070

    authors： Kumar A,Manimekalai MS,Balakrishna AM,Priya R,Biuković G,Jeyakanthan J,Grüber G

    更新日期：2010-09-03 00:00:00

  • The entericidin locus of Escherichia coli and its implications for programmed bacterial cell death.

    abstract：:Antidote/toxin gene pairs known as "addiction modules" can maintain plasmids in bacterial populations by means of post-segregational killing. However, several chromosome-encoded addiction modules may provide an entirely distinct function in the programmed cell death of moribund subpopulations under starvation conditio...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1998.1894

    authors： Bishop RE,Leskiw BK,Hodges RS,Kay CM,Weiner JH

    更新日期：1998-07-24 00:00:00

  • Derepression of SaPIbov1 Is Independent of φNM1 Type 2 dUTPase Activity and Is Inhibited by dUTP and dUMP.

    abstract：:Staphylococcus aureus is an opportunistic human pathogen able to transfer virulence genes to other cells through the mobilization of S. aureus pathogenicity islands (SaPIs). SaPIs are derepressed and packaged into phage-like transducing particles by helper phages like 80α or φNM1. Phages 80α and φNM1 encode structural...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2017.04.001

    authors： Hill RLL,Vlach J,Parker LK,Christie GE,Saad JS,Dokland T

    更新日期：2017-05-19 00:00:00

  • The 0.93A crystal structure of sphericase: a calcium-loaded serine protease from Bacillus sphaericus.

    abstract：:We have previously isolated sphericase (Sph), an extracellular mesophilic serine protease produced by Bacillus sphaericus. The Sph amino acid sequence is highly homologous to two cold-adapted subtilisins from Antarctic bacilli S39 and S41 (76% and 74% identity, respectively). Sph is calcium-dependent, 310 amino acid r...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2003.07.011

    authors： Almog O,González A,Klein D,Greenblatt HM,Braun S,Shoham G

    更新日期：2003-10-03 00:00:00

  • Structural basis of HIV-1 activation by NF-kappaB--a higher-order complex of p50:RelA bound to the HIV-1 LTR.

    abstract：:The activation and latency of human immunodeficiency virus type 1 (HIV-1) are tightly controlled by the transcriptional activity of its long terminal repeat (LTR) region. The LTR is regulated by viral proteins as well as host factors, including the nuclear factor kappaB (NF-kappaB) that becomes activated in virus-infe...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2009.08.023

    authors： Stroud JC,Oltman A,Han A,Bates DL,Chen L

    更新日期：2009-10-16 00:00:00

  • Bile acid interactions with rabbit ileal lipid binding protein and an engineered helixless variant reveal novel ligand binding properties of a versatile beta-clam shell protein scaffold.

    abstract：:The intracellular ileal lipid binding proteins (ILBPs) are involved in the transport and enterohepatic circulation of bile acids. ILBPs from different species show high sequence and structural homology and have been shown to bind multiple bile acid ligands with differing degrees of selectivity and positive co-operativ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2007.06.024

    authors： Kouvatsos N,Thurston V,Ball K,Oldham NJ,Thomas NR,Searle MS

    更新日期：2007-08-31 00:00:00

  • The effect of chloramphenicol in intact erythroid cells.

    abstract：:Incubation of intact reticulocytes with chloramphenicol at concentrations over 10(-3) M caused inhibition of protein synthesis and conversion of polyribosomes to single ribosomes. At 3 x 10(-3) M-chloramphenicol these effects were completely reversible, but reversibility decreased as the chloramphenicol concentration ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/0022-2836(66)90026-x

    authors： Godchaux W 3rd,Herbert E

    更新日期：1966-11-28 00:00:00

  • Bacterial Signaling to the Nervous System through Toxins and Metabolites.

    abstract：:Mammalian hosts interface intimately with commensal and pathogenic bacteria. It is increasingly clear that molecular interactions between the nervous system and microbes contribute to health and disease. Both commensal and pathogenic bacteria are capable of producing molecules that act on neurons and affect essential ...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章,评审

    doi：10.1016/j.jmb.2016.12.023

    authors： Yang NJ,Chiu IM

    更新日期：2017-03-10 00:00:00

  • Characterization of preferred deoxyribonuclease I cleavage sites.

    abstract：:The preferred DNase I cleavage sites within the 160 bp tyrT DNA fragment were identified by studying the initial rate of cleavage of individual bonds. The results show that there is no correlation between the rate of cleavage and the identity of the dinucleotide sequence that is cleaved. Examination of the sequences s...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1994.1152

    authors： Herrera JE,Chaires JB

    更新日期：1994-02-18 00:00:00

  • RNA binding, packaging and polymerase activities of the different incomplete polymerase complex particles of dsRNA bacteriophage phi 6.

    abstract：:phi 6 is an enveloped dsRNA bacterial virus. Its segmented genome resides inside the virion associated polymerase complex which is formed by four proteins (P1, P2, P4 and P7) encoded by the viral L segment. Complete and incomplete polymerase complex particles can be produced using cDNA copies of this largest genome se...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1995.0317

    authors： Juuti JT,Bamford DH

    更新日期：1995-06-09 00:00:00

  • Cooperativity in the thermosome.

    abstract：:The thermosome from Thermoplasma acidophilum is a type II chaperonin composed of eight alpha and eight beta subunits. The genes encoding the two types of subunit were co-expressed in Escherichia coli and the alpha8/beta8 complex purified from the cell extract. The isolated complex showed steady-state ATPase properties...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2005.01.066

    authors： Bigotti MG,Clarke AR

    更新日期：2005-04-22 00:00:00

  • Optimal packaging of FIV genomic RNA depends upon a conserved long-range interaction and a palindromic sequence within gag.

    abstract：:The feline immunodeficiency virus (FIV) is a lentivirus that is related to human immunodeficiency virus (HIV), causing a similar pathology in cats. It is a potential small animal model for AIDS and the FIV-based vectors are also being pursued for human gene therapy. Previous studies have mapped the FIV packaging signa...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2010.08.019

    authors： Rizvi TA,Kenyon JC,Ali J,Aktar SJ,Phillip PS,Ghazawi A,Mustafa F,Lever AML

    更新日期：2010-10-15 00:00:00

  • Fast Translation within the First 45 Codons Decreases mRNA Stability and Increases Premature Transcription Termination in E. coli.

    abstract：:We show here that the specific use of fast or slowly translated codons in the early coding region of a gene may influence both the mRNA stability and premature transcription termination. We first inserted a pair of nearly identical 42-base-pair (bp)-long sequences into codon 3 of the Escherichia coli lacZ gene. The on...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2019.01.026

    authors： Pedersen S,Terkelsen TB,Eriksen M,Hauge MK,Lund CC,Sneppen K,Mitarai N

    更新日期：2019-03-15 00:00:00

  • Structure of a foreign peptide displayed on the surface of bacteriophage M13.

    abstract：:The use of filamentous bacteriophage M13 as a vehicle for display of foreign peptides and proteins provides a means for the construction of therapeutic, diagnostic and technological tools of broad utility. The usefulness of this technology is dependent on the ability of an inserted peptide to act as a ligand when fuse...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1006/jmbi.1994.1489

    authors： Kishchenko G,Batliwala H,Makowski L

    更新日期：1994-08-12 00:00:00

  • Interactions of the natural product kendomycin and the 20S proteasome.

    abstract：:Natural products are a valuable source for novel lead structures in drug discovery, but for the majority of isolated bioactive compounds, the cellular targets are unknown. The structurally unique ansa-polyketide kendomycin (KM) was reported to exert its potent cytotoxic effects via impairment of the ubiquitin proteaso...

    journal_title：Journal of molecular biology

    pub_type： 杂志文章

    doi：10.1016/j.jmb.2014.06.019

    authors： Beck P,Heinemeyer W,Späth AL,Elnakady Y,Müller R,Groll M

    更新日期：2014-09-09 00:00:00