The algorithm of estimation of the Km values for primers of various structure and length in the polymerization reaction catalyzed by Klenow fragment of DNA polymerase I from E. coli.

Abstract:

:DNA synthesis at primers d(pT)n, d(pA)n, d(pC)n, and d(pG)n in the presence of corresponding complementary templates and at hetero-oligoprimers complementary to M13 phage DNA was investigated. The values of both -log Km and log Vmax increased linearly if homo-oligoprimers contained less than 10 nucleotides. The lengthening of d(pT)n and d(pA)n primers by one mononucleotide unit (n = 1-10) resulted in the 1.82-fold decrease of the Km values. The incremental decreases of Km for d(pC)n and d(pG)n were equal to about 2.46. The enhancement of the homo- and hetero-oligonucleotide primers' affinity to the enzyme due to one Watson-Crick hydrogen bond between complementary template and primer is about 1.35 times. This allows to calculate the Km values for primers of various structure and length up to 10 units. The objective laws of the Km and Vmax values changes for primers containing more than 10 nucleotides were analyzed.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Nevinsky GA,Nemudraya AV,Levina AS,Khomov VV

doi

10.1016/0014-5793(89)81642-4

subject

Has Abstract

pub_date

1989-11-20 00:00:00

pages

166-70

issue

1

eissn

0014-5793

issn

1873-3468

pii

0014-5793(89)81642-4

journal_volume

258

pub_type

杂志文章
  • Human S100A9 potentiates IL-8 production in response to GM-CSF or fMLP via activation of a different set of transcription factors in neutrophils.

    abstract::Inflammation is highly regulated by various agents. Unexpectedly, we report here that the damage-associated molecular pattern S100A9 protein, a potent neutrophil activator and inducer of cytokine production in monocytes, is not a direct activator of cytokine production in human neutrophils. However, S100A9 primed IL-8...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2014.04.027

    authors: Simard JC,Noël C,Tessier PA,Girard D

    更新日期:2014-06-13 00:00:00

  • Asparaginyl-tRNA synthetase from the Escherichia coli temperature-sensitive strain HO202. A proline replacement in motif 2 is responsible for a large increase in Km for asparagine and ATP.

    abstract::The Escherichia coli K12 mutant gene, asnS40, coding for asparaginyl-tRNA synthetase (AsnRS) in the temperature-sensitive strain HO202, was isolated from genomic DNA using the Polymerase Chain Reaction. DNA sequencing revealed that the mutant enzyme differs from the wild-type AsnRS by two amino acids, but only the P23...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(92)80106-q

    authors: Madern D,Anselme J,Härtlein M

    更新日期:1992-03-24 00:00:00

  • Transcriptional enhancer factor 1 (TEF-1/TEAD1) mediates activation of IFITM3 gene by BRGl.

    abstract::The interferon inducible transmembrane (IFITM) proteins mediate several cellular processes such as homotypic cell adhesion functions of interferons (IFNs) and cellular anti-proliferative activities. We show that the BAF complex-mediated induction of IFITM3 is dependent on binding of the transcriptional enhancer factor...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2007.12.033

    authors: Cuddapah S,Cui K,Zhao K

    更新日期:2008-01-23 00:00:00

  • Evidence from peptide mapping for a human renin zymogen.

    abstract::Data obtained from peptide mapping of the active and inactive forms of human renin show that there are extensive regions of common sequence in the two forms of the enzyme, and are consistent with the hypothesis that inactive renin is a renin zymogen. ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(84)80784-x

    authors: McIntyre GD,Chaudhuri S,Coggins JR

    更新日期:1984-10-01 00:00:00

  • Visualisation by electron microscopy of the unique part of the cytoplasmic domain of a desmoglein, a cadherin-like protein of the desmosome type of cell junction.

    abstract::Part of the cytoplasmic domain of a human desmoglein, Dsg1, a cadherin-like protein found in desmosomes of epithelial cells, has been visualised by electron microscopy. The cloned fragment contains five repeats of a 29 +/- 4 residue sequence unique to desmogleins, followed by a glycine-rich region. In rotary shadowed ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(94)01049-8

    authors: Rutman AJ,Buxton RS,Burdett ID

    更新日期:1994-10-17 00:00:00

  • Robustness, evolvability, and neutrality.

    abstract::Biological systems, from macromolecules to whole organisms, are robust if they continue to function, survive, or reproduce when faced with mutations, environmental change, and internal noise. I focus here on biological systems that are robust to mutations and ask whether such systems are more or less evolvable, in the...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2005.01.063

    authors: Wagner A

    更新日期:2005-03-21 00:00:00

  • Downregulation of collagen synthesis in fibroblasts within three-dimensional collagen lattices involves transcriptional and posttranscriptional mechanisms.

    abstract::Culturing human fibroblasts in a three-dimensional collagen matrix leads to a reduction of collagen I by more than 90%, both on the level of mRNA steady-state as well as protein. In order to differentiate changes in de novo transcription and posttranscriptional control, nuclear run on assays and pulse/chase experiment...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(93)80006-g

    authors: Eckes B,Mauch C,Hüppe G,Krieg T

    更新日期:1993-03-01 00:00:00

  • Role of thioredoxin-1 in apoptosis induction by alpha-tocopheryl succinate and TNF-related apoptosis-inducing ligand in mesothelioma cells.

    abstract::Malignant mesothelioma (MM) is a fatal type of cancer. We studied the role of the redox-active protein thioredoxin-1 (Trx-1) in apoptosis induced in MM cells and their non-malignant counterparts (Met-5A) by alpha-tocopheryl succinate (alpha-TOS) and TNF-related apoptosis-inducing ligand (TRAIL). MM cells were suscepti...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2006.04.019

    authors: Freeman RE,Neuzil J

    更新日期:2006-05-15 00:00:00

  • Presynaptic kainate receptor-mediated facilitation of glutamate release involves Ca2+-calmodulin and PKA in cerebrocortical synaptosomes.

    abstract::We have explored the mechanisms involved in the facilitation of glutamate release mediated by the activation of kainate receptors (KARs) in the cortex using isolated nerve terminals (synaptosomes). Kainate (KA) produced an increase on glutamate release at 100 μM. The effect of KA was antagonized by NBQX (with AMPA rec...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2013.01.071

    authors: Rodríguez-Moreno A,Sihra TS

    更新日期:2013-03-18 00:00:00

  • Apolipoprotein E associated with reconstituted high-density lipoprotein-like particles is protected from aggregation.

    abstract::Apolipoprotein E (APOE) genotype determines Alzheimer's disease (AD) susceptibility, with the APOE ε4 allele being an established risk factor for late-onset AD. The ApoE lipidation status has been reported to impact amyloid-beta (Aβ) peptide metabolism. The details of how lipidation affects ApoE behavior remain to be ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1002/1873-3468.13428

    authors: Hubin E,Verghese PB,van Nuland N,Broersen K

    更新日期:2019-06-01 00:00:00

  • Preparation of peptides which mimic glycosphingolipids by using phage peptide library and their modulation on beta-galactosidase activity.

    abstract::We describe the use of a phage-displayed random pentadecamer peptide library for searching glycosphingolipid mimicking peptides. Two phage clones (AD-1 and AD-2) were selected by biopanning using monoclonal antibody AD117m, directed to lactotetraosylceramide (Lc4Cer). The amino acid sequences of the selected clones sh...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(97)01386-0

    authors: Taki T,Ishikawa D,Hamasaki H,Handa S

    更新日期:1997-11-24 00:00:00

  • Involvement of AMP-activated protein kinase in fat depot-specific metabolic changes during starvation.

    abstract::The mechanisms controlling fat depot-specific metabolism are poorly understood. During starvation of mice, downregulation of lipogenic genes, suppression of fatty acid synthesis, and increases in lipid oxidation were all more pronounced in epididymal than in subcutaneous fat. In epididymal fat, relatively strong upreg...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2005.09.078

    authors: Sponarova J,Mustard KJ,Horakova O,Flachs P,Rossmeisl M,Brauner P,Bardova K,Thomason-Hughes M,Braunerova R,Janovska P,Hardie DG,Kopecky J

    更新日期:2005-11-07 00:00:00

  • Activation of a capacitative Ca(2+) entry pathway by store depletion in cultured hippocampal neurones.

    abstract::Intracellular Ca(2+) ([Ca(2+)](i)) changes were measured in cell bodies of cultured rat hippocampal neurones with the fluorescent indicator Fluo-3. In the absence of external Ca(2+), the cholinergic agonist carbachol (200 microM) and the sarcoendoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (0.4 microM) both t...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(00)01340-5

    authors: Bouron A

    更新日期:2000-03-31 00:00:00

  • Discovery of the catalytic function of a putative 2-oxoacid dehydrogenase multienzyme complex in the thermophilic archaeon Thermoplasma acidophilum.

    abstract::Those aerobic archaea whose genomes have been sequenced possess a single 4-gene operon that, by sequence comparisons with Bacteria and Eukarya, appears to encode the three component enzymes of a 2-oxoacid dehydrogenase multienzyme complex. However, no catalytic activity of any such complex has ever been detected in th...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2004.10.058

    authors: Heath C,Jeffries AC,Hough DW,Danson MJ

    更新日期:2004-11-19 00:00:00

  • Increased muscarinic receptor binding in heart membranes by an inhibitor of protein kinase C.

    abstract::The number of muscarinic acetylcholine receptors (MAChRs), as detected by binding of [3H]quinuclidinyl benzilate (QNB), was investigated under conditions which promote protein phosphorylation. Incubation of a crude heart membrane preparation in the presence of ATP/Mg2+ reduced MAChR number by 50%. Incubation with poly...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(88)81010-x

    authors: Creazzo TL,Wrenn RW

    更新日期:1988-12-19 00:00:00

  • Transcriptional changes in response to growth of Arabidopsis in high external calcium.

    abstract::Transcriptional responses to growth in high environmental calcium concentrations were characterized and compared between wild-type and mutant Arabidopsis plants containing a knockout mutation in the gene encoding a cyclic nucleotide-gated channel (CNGC2). We show that the transcriptional profile of cngc2 plants grown ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2008.02.043

    authors: Chan CW,Wohlbach DJ,Rodesch MJ,Sussman MR

    更新日期:2008-03-19 00:00:00

  • Actin microridges characterized by laser scanning confocal and atomic force microscopy.

    abstract::We have characterized the cell surface of zebrafish stratified epithelium using a combined approach of light and atomic force microscopy under conditions which simulate wound healing. Microridges rise on average 100 nm above the surface of living epithelial cells, which correlate to bundles of cytochalasin B-insensiti...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2005.02.049

    authors: Sharma A,Anderson KI,Müller DJ

    更新日期:2005-03-28 00:00:00

  • Reactivity of glutaredoxins 1, 2 and 3 from Escherichia coli and protein disulfide isomerase towards glutathionyl-mixed disulfides in ribonuclease A.

    abstract::We have examined the activity of protein disulfide isomerase (PDI) and glutaredoxin (Grx) 1, 2 and 3 from Escherichia coli to catalyze the cleavage of glutathionylated ribonuclease A (RNase-SG) by 1 mM GSH to yield reduced RNase. Apparent Km values for RNase-SG were similar, 2-10 microM, for Grx 1, 3 and PDI but Grx I...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(98)01698-6

    authors: Lundström-Ljung J,Vlamis-Gardikas A,Aslund F,Holmgren A

    更新日期:1999-01-25 00:00:00

  • Maturation and degradation of beta-galactosidase in the post-Golgi compartment are regulated by cathepsin B and a non-cysteine protease.

    abstract::Lysosomal beta-galactosidase precursor is processed to a mature form and associated with protective protein in lysosomes. In this study we used two cysteine protease proinhibitors, E64-d for cathepsins B, S, H, and L, and CA074Me for cathepsin B. They are converted intracellularly to active forms, E-64c and CA074, res...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(97)01461-0

    authors: Okamura-Oho Y,Zhang S,Callahan JW,Murata M,Oshima A,Suzuki Y

    更新日期:1997-12-15 00:00:00

  • Reversible inactivation of endothelial nitric oxide synthase by NG-nitro-L-arginine.

    abstract::NG-Methyl-L-arginine (L-NMA) and NG-nitro-L-arginine (L-NNA) inhibited NO-induced cGMP accumulation in porcine aortic endothelial cells with half-maximally effective concentrations of 15 and 3.4 microM, respectively. The effects of both compounds were reversible, but the L-NNA-induced inhibition was only reversed by w...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(93)80405-j

    authors: Mayer B,Schmid M,Klatt P,Schmidt K

    更新日期:1993-10-25 00:00:00

  • Acid phospholipase A activities in rat hepatocytes.

    abstract::Cultured rat hepatocytes exhibit acid phospholipase A activity. On the basis of product formation from stereospecifically radiolabeled phosphatidylethanolamine substrates, phospholipases A1 and A2 have been identified with optimal activities at pH 4.5. According to subcellular fractionation studies, the acid phospholi...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(88)80062-0

    authors: Kunze H,Löffler BM,Schmidt M

    更新日期:1988-08-29 00:00:00

  • Mechanism for the peroxynitrite scavenging activity by anthocyanins.

    abstract::We show that anthocyanins can function as potent inhibitors of the formation of nitrated tyrosine in vitro, and clarified how pelargonidin (Pel), which has a mono-hydroxyl group on the B-ring, can scavenge peroxynitrite (ONOO(-)) by detection of the reaction products. Pel was reacted with ONOO(-), then the reaction mi...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(00)02150-5

    authors: Tsuda T,Kato Y,Osawa T

    更新日期:2000-11-10 00:00:00

  • Constructing an efficient trans-acting genomic HDV ribozyme.

    abstract::We have engineered a genomic HDV ribozyme to construct several trans-acting ribozymes for use in trans to cleave target RNAs. Among the 10 different combinations attempted, only HDV88-Trans had cleavage activity on the 13-nucleotide substrate, R13, in vitro. To improve the cleavage efficiency, at least in vitro, of th...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(96)00941-6

    authors: Kawakami J,Yuda K,Suh YA,Kumar PK,Nishikawa F,Maeda H,Taira K,Ohtsuka E,Nishikawa S

    更新日期:1996-09-30 00:00:00

  • Mitochondrial Isa2p plays a crucial role in the maturation of cellular iron-sulfur proteins.

    abstract::The assembly of iron-sulfur (Fe/S) clusters in a living cell is mediated by a complex machinery which, in eukaryotes, is localised within mitochondria. Here, we report on a new component of this machinery, the protein Isa2p of the yeast Saccharomyces cerevisiae. The protein shares sequence similarity with yeast Isa1p ...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/s0014-5793(00)01711-7

    authors: Pelzer W,Mühlenhoff U,Diekert K,Siegmund K,Kispal G,Lill R

    更新日期:2000-07-07 00:00:00

  • NPC1L1 and cholesterol transport.

    abstract::The polytopic transmembrane protein, Niemann-Pick C1-Like 1 (NPC1L1), is enriched in the apical membrane of small intestine absorptive enterocytes where it mediates extracellular sterol transport across the brush border membrane. It is essential for intestinal sterol absorption and is the molecular target of ezetimibe...

    journal_title:FEBS letters

    pub_type: 杂志文章,评审

    doi:10.1016/j.febslet.2010.03.030

    authors: Betters JL,Yu L

    更新日期:2010-07-02 00:00:00

  • Isolation and sequence analysis of a cDNA clone encoding the entire catalytic subunit of phosphorylase kinase.

    abstract::Synthetic oligonucleotides have been used to isolate a 1.85 kb clone containing the full length coding sequence for the catalytic subunit of rabbit skeletal muscle phosphorylase kinase from a cDNA library constructed in lambda gt10. Sequence analysis of the clone predicted an amino acid sequence in agreement with a pu...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(87)80871-2

    authors: da Cruz e Silva EF,Cohen PT

    更新日期:1987-08-10 00:00:00

  • Endothelin-1 induces hypoxia inducible factor 1α expression in pulmonary artery smooth muscle cells.

    abstract::Endothelin-1 (ET-1) dose-dependently increased HIF1α expression in pulmonary artery smooth muscle cells (PASMCs). Inhibition of protein synthesis did not affect ET-1-induced HIF1α expression. The maximum effect of ET-1 was similar to that caused by proteasome inhibitor MG132. Further study indicates that ET-1 also dos...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.08.036

    authors: Li M,Liu Y,Jin F,Sun X,Li Z,Liu Y,Fang P,Shi H,Jiang X

    更新日期:2012-11-02 00:00:00

  • Structural and functional analysis of Trichoderma reesei endoglucanase I expressed in yeast Saccharomyces cerevisiae.

    abstract::The function of the domains of Trichoderma reesei endoglucanase I (EGI) has been studied. Truncated EGI proteins were expressed from the 3'-end deleted cDNAs in the yeast Saccharomyces cerevisiae under the control of the ADC1 expression cassette. EGI protein was detected by monoclonal antibody EI-2 and EGI activity as...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(91)81100-m

    authors: Aho S

    更新日期:1991-10-07 00:00:00

  • Transient small molecule interactions kinetically modulate amyloid β peptide self-assembly.

    abstract::Small organic molecules, like Congo red and lacmoid, have been shown to modulate the self-assembly of the amyloid β peptide (Aβ). Here, we show that Aβ forms NMR invisible non-toxic co-aggregates together with lacmoid as well as Congo red. We find that the interaction involves two distinct kinetic processes and at eve...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/j.febslet.2012.09.035

    authors: Abelein A,Lang L,Lendel C,Gräslund A,Danielsson J

    更新日期:2012-11-16 00:00:00

  • Formation of free sphingosine and ceramide from exogenous ganglioside GM1 by cerebellar granule cells in culture.

    abstract::Cerebellar granule cells differentiated in culture were incubated with ganglioside [3H-Sph]GM1 in order to have it inserted into the plasma membrane and metabolized. Among the formed metabolites radioactive sphingosine and ceramide were identified. [3H]Ceramide started to be measurable after 10 min of incubation (puls...

    journal_title:FEBS letters

    pub_type: 杂志文章

    doi:10.1016/0014-5793(92)80193-k

    authors: Riboni L,Bassi R,Sonnino S,Tettamanti G

    更新日期:1992-03-30 00:00:00