Abstract:
:5,6-Dihydrothymidine 5'-triphosphate (DHdTTP) was synthesized by catalytic hydrogenation of thymidine 5'-triphosphate (dTTP). Thymidine glycol 5'-triphosphate (dTTP-GLY) was prepared by bromination of dTTP followed by treatment with Ag2O. The modified nucleotides were extensively purified by anion-exchange high-performance liquid chromatography (HPLC). Alkaline phosphatase digestion of DHdTTP and dTTP-GLY gave the expected products (5,6-dihydrothymidine and cis-thymidine glycol), the identities of which were confirmed by reverse-phase HPLC using authentic markers. HPLC analysis of the alkaline phosphatase digested DHdTTP revealed that DHdTTP was a mixture of C5 diastereoisomers [(5S)- and (5R)-DHdTTP]. Despite the significant distortion of the pyrimidine ring in DHdTTP, it was incorporated in place of dTTP during primer elongation catalyzed by Escherichia coli DNA polymerase I Klenow fragment. The rate of incorporation of DHdTTP was about 10-25-fold lower than that of dTTP. On the other hand, dTTP-GLY, which also has a distorted pyrimidine ring, did not replace dTTP, and no elongation of the primer was observed. In order to study the preference of incorporation of the diastereoisomers of DHdTTP into DNA, salmon testes DNA, activated by exonuclease III, was used as a template for DNA polymerase I Klenow fragment in the presence of [3H]DHdTTP (S and R mixture) and normal nucleotides. After enzymatic digestion of the DNA to nucleosides, the products were analyzed by HPLC. The ratio of the isomers incorporated into DNA (S:R = 73.27) was virtually the same as that of the [3H]DHdTTP substrates (S:R = 79.21).(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Ide H,Melamede RJ,Wallace SSdoi
10.1021/bi00377a042subject
Has Abstractpub_date
1987-02-10 00:00:00pages
964-9issue
3eissn
0006-2960issn
1520-4995journal_volume
26pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Electrophilic halogenating agents, including hypohalous acids and haloamines, oxidize free methionine and the N-terminal methionines of peptides and proteins (e.g., Met-1 of anti-inflammatory peptide 1 and ubiquitin) to produce dehydromethionine (a five-membered isothiazolidinium heterocycle). Amide derivatives of met...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901343d
更新日期:2009-11-24 00:00:00
abstract::Attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy has been performed on samples of bovine cytochrome c oxidase that have been deposited as a thin film on the surface of a silicon microprism. The technique has several advantages over transmission methods in terms of amount of mat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0109717
更新日期:2002-01-22 00:00:00
abstract::Synthetic nonviral vector systems are attractive because of their apparent simplicity of preparation and use. However, there are many barriers to success at the moment, including the formulation of uniform and reproducible particles of transfection competent, condensed nucleic acids such as plasmid DNA (pDNA). For thi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034325e
更新日期:2003-09-09 00:00:00
abstract::A general understanding of how cytochromes evolve within a fixed structure to optimize redox potential for specific bioenergetic processes does not exist. Toward this end, a library approach is used to investigate the range and distribution of redox potential which occurs when all sequence space available through muta...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0001675
更新日期:2000-05-23 00:00:00
abstract::N-[[(Iodoacetyl)amino]ethyl]-5-naphthylamine-1-sulfonic acid (IAEDANS) is a fluorescent reagent which reacts covalently with the free thiol groups of proteins. When the reagent is reacted with the Escherichia coli ribosome under mild conditions, gel electrophoresis shows modification of predominantly two proteins, S18...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00346a042
更新日期:1985-12-03 00:00:00
abstract::The carboxymethylation of RNase T1 at the gamma-carboxyl group of Glu58 leads to a complete loss of the enzymatic activity while it retains substrate-binding ability. Accompanying the carboxymethylation, RNase T1 undergoes a remarkable thermal stabilization of 9 degrees C in the melting temperature (Tm). In order to c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960493d
更新日期:1996-06-25 00:00:00
abstract::Double strand breaks in DNA can be quantified down to very low frequencies (a few per Gigabase pair) in nanogram quantities of nonradioactive, genomic DNA by dispersing the DNAs on electrophoretic gels, digitizing them by quantitative electronic imaging, and calculating the DNA lengths by number average length analysi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0205505
更新日期:2003-03-25 00:00:00
abstract::The pathogenic bacteria Pseudomonas aeruginosa and Bordetella pertussis contain in their outer membranes the rare sugar 2,3-diacetamido-2,3-dideoxy-d-mannuronic acid. Five enzymes are required for the biosynthesis of this sugar starting from UDP-N-acetylglucosamine. One of these, referred to as WlbB, is an N-acetyltra...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005738
更新日期:2010-06-08 00:00:00
abstract::In the present study hydrostatic pressure was applied upon both skeletal myosin rod molecules and rod minifilaments to learn more of the intra- and intermolecular interaction behavior of myosin. Applied pressure disassembled the rod minifilaments into individual rod molecules and dissociated each myosin rod molecule i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00184a028
更新日期:1994-05-10 00:00:00
abstract::To understand the mechanism by which annexin I induces membrane aggregation, a comprehensive mutagenesis of all six Ca2+-binding sites was performed. When the cap residues of type II Ca2+-binding sites were systematically mutated to Ala, a type II site in domain II was shown to be essential for Ca2+-dependent vesicle ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980479w
更新日期:1998-07-14 00:00:00
abstract::The clam Scapharca inaequivalvis possesses two cooperative oxygen binding hemoglobins in its red cells: a homodimeric HbI and a heterotetrameric A2B2 HbII. Each AB dimeric half of HbII is assembled in a manner very similar to that of the well-studied HbI. This study presents crystal structures of HbII along with oxyge...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi301620x
更新日期:2013-03-26 00:00:00
abstract::Hepatic mixed-function oxidase metabolism of the ubiquitous pollutant polychlorinated biphenyls (PCBs) is implicated in their toxification and detoxification. We used dichlorobiphenyls (DCBs) as models to investigate the effect of the chloro substituent sites on this metabolism experimentally and by molecular orbital ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00529a009
更新日期:1981-12-22 00:00:00
abstract::Octaprenyl pyrophosphate synthase (OPPs) catalyzes consecutive condensation reactions of farnesyl pyrophosphate (FPP) with five molecules of isopentenyl pyrophosphate (IPP) to generate C(40) octaprenyl pyrophosphate (OPP) which constitutes the side chain of menaquinone. We have previously reported the X-ray structure ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi036336d
更新日期:2004-06-22 00:00:00
abstract::NADP(H) binding is essential for fast electron transfer through the flavoprotein domain of the fusion protein P450BM3. Here we characterize the interaction of NADP(H) with the oxidized and partially reduced enzyme and the effect of this interaction on the redox properties of flavin cofactors and electron transfer. Mea...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001068u
更新日期:2000-10-17 00:00:00
abstract::Apo-electron-transferring flavoprotein from pig kidney (apo-ETF) has been prepared by an acid ammonium sulfate procedure and reconstituted with FAD analogues to probe the flavin binding site. The 8-position of the bound flavin is accessible to solvent as judged by the reaction of 8-Cl-FAD-ETF with sodium sulfide and t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00370a050
更新日期:1986-11-04 00:00:00
abstract::Endothelium-derived epoxyeicosatrienoic acids (EETs) relax vascular smooth muscle by activating potassium channels and causing membrane hyperpolarization. Recent evidence suggests that EETs act via a membrane binding site or receptor. To further characterize this binding site or receptor, we synthesized 20-iodo-14,15-...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi102070w
更新日期:2011-05-10 00:00:00
abstract::A series of 24-homologated 1,25-dihydroxyvitamin D3 compounds have been chemically synthesized and studied with regard to their activity in inducing differentiation of human promyelocyte HL-60 cells to monocytes and in calcium mobilizing activity in vitamin D deficient rats. Homologation of 1,25-dihydroxyvitamin D3 or...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00453a026
更新日期:1990-01-09 00:00:00
abstract::Inhibition of gamma-aminobutyric acid aminotransferase (GABA-AT) increases the concentration of GABA, an inhibitory neurotransmitter in human brain, which could have therapeutic applications for a variety of neurological diseases, including epilepsy. On the basis of studies of several previously synthesized conformati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061592m
更新日期:2006-12-05 00:00:00
abstract::The structure and dynamic properties of the C-terminal region of the human class alpha glutathione transferase A1-1 have been investigated with high-resolution NMR methods. On the basis of crystallographic and fluorescence measurements, this 13-residue segment of the enzyme is presumed to be disordered in the unligand...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0363329
更新日期:2004-06-15 00:00:00
abstract::We report nuclear magnetic resonance (NMR) data indicating that the Rieske protein from the cytochrome bc complex of Thermus thermophilus (TtRp) undergoes modest redox-state-dependent and ligand-dependent conformational changes. To test models concerning the mechanism by which TtRp transfers between different sites on...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400296c
更新日期:2013-04-30 00:00:00
abstract::First identified on the surface of lymphoids as a type II transmembrane protein, CD38 has now been established to have dual functions not only as a receptor but also as a multifunctional enzyme,catalyzing the synthesis of and hydrolysis of a general calcium messenger molecule, cyclic ADP-ribose(cADPR). The receptorial...
journal_title:Biochemistry
pub_type: 杂志文章
doi:
更新日期:2008-12-30 00:00:00
abstract::The chromatin structure encompassing the lysozyme gene domain in hen oviduct nuclei was studied by measuring the partitioning of coding and flanking sequences during chromatin fractionation and by analyzing the nucleosome repeat in response to micrococcal nuclease digestion. Following micrococcal nuclease digestion, n...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00350a033
更新日期:1986-01-28 00:00:00
abstract::The fluorescent nucleotide analogue of ADP, 1,N6-ethenoadenosine diphosphate (epsilon ADP), has been used to probe the active site of myosin subfragment 1 (SF1). The Mg complex of ADP was shown to be trapped stoichiometrically at the active site by a variety of thiol cross-linking agents having sulfur to sulfur spanni...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00312a029
更新日期:1984-08-14 00:00:00
abstract::The kinetics and thermodynamics of Al(III) exchange between aluminum citrate (AlL) and human serum transferrin were investigated in the 7.2-8.9 pH range. The C-site of human serum apotransferrin in interaction with bicarbonate removes Al(III) from Al citrate with an exchange equilibrium constant K1 = (2.0 +/- 0.6) x 1...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi020627p
更新日期:2003-03-18 00:00:00
abstract::The interaction of aminooxy compounds such as aminooxyacetate (AAA), L-canaline, and hydroxylamine with sheep liver serine hydroxymethyltransferase (EC 2.1.2.1) was studied by absorption spectra and stopped-flow spectrophotometry and compared with the unique feature of interaction of O-amino-D-serine (OADS) with the e...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00451a010
更新日期:1989-12-12 00:00:00
abstract::Locusta migratoria apolipophorin III (apoLp-III) is a helix bundle exchangeable apolipoprotein that reversibly binds to lipoprotein surfaces. Structural reorganization of its five amphipathic alpha-helices enables the transition from the lipid-free to lipid-bound state. ApoLp-III-induced transformation of dimyristoylp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010410f
更新日期:2001-06-26 00:00:00
abstract::Calmodulin and troponin C exhibit calcium-dependent binding of 1 mol/mol of dynorphin. The dissociation constants of the complexes, determined in 0.20 N KC1-1.0 mM CaCI2, pH 7.3, are 0.6 microM for calmodulin, 2.4 microM for rabbit fast skeletal muscle troponin C, and 9 microM for bovine heart troponin C. Experiments ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00306a016
更新日期:1984-05-22 00:00:00
abstract::Organisms rely on regulation at the molecular level, such as the allosteric regulation of hemoglobin (Hb) function by anions, to meet challenges presented by changing environmental and physiological conditions. A comparison of the effects of anions on oxygenation, oxidation, and sulfhydryl reactivity of Hb leads us to...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971574s
更新日期:1998-01-13 00:00:00
abstract::Ala-based peptides form marginally stable helices at low temperature and are conventionally considered as mixtures of alpha-helix and random coil. However, recent work with doubly spin-labeled peptides suggests that short 16-residue sequences contain a significant fraction of 3(10)-helix near the N-terminus (positions...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00096a003
更新日期:1993-11-16 00:00:00
abstract::Human alpha-lactalbumin (alpha-LA), a 123-residue calcium-binding protein, has been studied using (15)N NMR relaxation methods in order to characterize backbone dynamics of the native state at the level of individual residues. Relaxation data were collected at three magnetic field strengths and analyzed using the mode...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi900023m
更新日期:2009-05-19 00:00:00