Abstract:
:Human S-adenosylmethionine decarboxylase (AdoMetDC) was expressed in high yield in Escherichia coli using the pIN-III(lppP-5) expression vector and purified to apparent homogeneity using affinity chromatography on methylglyoxal bis(guanylhydrazone)-Sepharose. The inactivation of the purified enzyme by 5'-deoxy-5'-[(3-hydrazinopropyl)methylamino]adenosine (MHZPA) was accompanied by an increase in absorbance at 260 nm of the large subunit. This increase was equivalent to the addition of 1 molecule of MHZPA. After digestion with the protease Lys-C, a peptide that contained the bound MHZPA was isolated and found to have the amino acid composition consistent with that expected from the amino terminus of the large subunit. These results indicate that MHZPA inactivates AdoMetDC by forming a hydrazone derivative at the pyruvate prosthetic group. Inactivation of AdoMetDC by 5'-([(Z)-4-amino-2-butenyl]methylamino]-5'-deoxyadenosine (AbeAdo) led to the appearance of a new peptide peak in the Lys-C protease digest. This peptide had the sequence ASMFVSK. This agrees with the expected sequence from the amino terminus, which is pyruvoyl-SMFVSK, with the exception that the pyruvate has been converted to alanine. Direct gas-phase sequencing of the large subunit of the enzyme also indicated the presence of alanine at the amino terminus after inactivation with AbeAdo. These results indicate that this inhibitor leads to transamination of the pyruvate prosthetic group. Since the pyruvate is covalently linked to the protein, its replacement by alanine leads to an irreversible inactivation of AdoMetDC.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Shantz LM,Stanley BA,Secrist JA 3rd,Pegg AEdoi
10.1021/bi00144a027subject
Has Abstractpub_date
1992-07-28 00:00:00pages
6848-55issue
29eissn
0006-2960issn
1520-4995journal_volume
31pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Human prolactin (hPRL) binds two human prolactin receptor molecules, creating active heterotrimeric complexes. Receptors bind dissimilar hormone surfaces termed site 1 and site 2 in an obligate ordered process. We sought to map the functional epitopes in site 1 of hPRL. Extensive alanine mutagenesis (102 of the 199 re...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi101838s
更新日期:2011-03-01 00:00:00
abstract::Bacterial L-asparaginases, enzymes that catalyze the hydrolysis of L-asparagine to aspartic acid, have been used for over 30 years as therapeutic agents in the treatment of acute childhood lymphoblastic leukemia. Other substrates of asparaginases include L-glutamine, D-asparagine, and succinic acid monoamide. In this ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0029595
更新日期:2001-05-15 00:00:00
abstract::After borohydride reduction, carboxymethylation, and tryptic digestion of the holoenzyme of pig heart alanine aminotransferase, a single icosapeptide containing the N6-(phosphopyridoxyl)lysine residue was isolated by a combination of gel filtration and ion-exchange chromatogrpahy. Its primary structure was determined ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00581a015
更新日期:1979-07-10 00:00:00
abstract::c-Myc is a member of a family of sequence specific-DNA binding proteins that are thought to regulate the transcription of genes involved in normal cell growth, differentiation, and apoptosis. In order to understand how human c-myc functions as a transcription factor, we have studied the mechanism of action and structu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960793v
更新日期:1996-07-23 00:00:00
abstract::Since a major function of RNA polymerase must be to bring together substrates in the optimal configuration for internucleotide bond formation, studies have been undertaken to understand the geometrical relationship of the two substrates. A model has been constructed for the geometry of interaction of two ATP molecules...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00477a017
更新日期:1990-06-26 00:00:00
abstract::Proteases specific for cleavage after acidic residues have been implicated in several disease states, including epidermolysis, inflammation, and viral processing. A serine protease with specificity toward glutamic acid substrates (Glu-SGP) has been crystallized in the presence of a tetrapeptide ligand and its structur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00094a001
更新日期:1993-11-02 00:00:00
abstract::A recombinant system was developed for generation of steroid-receptor complexes in vitro. The DNA- and steroid-binding domains of the rat mineralocorticoid receptor were expressed in Escherichia coli as a fusion protein with glutathione S-transferase. The identity of the expressed recombinant protein was confirmed by ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00084a028
更新日期:1993-08-24 00:00:00
abstract::[1-Penicillamine,2-leucine]oxytocin is a conformationally restricted analogue of oxytoxin in which the half-cystine-1 and tyrosine-2 residues of the native hormone are replaced by half-penicillamine (beta, beta-dimethyl-half-cystine) and leucine, respectively. This analogue is a surprisingly potent oxytocin antagonist...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00513a018
更新日期:1981-05-12 00:00:00
abstract::The enzymes in the alpha-ketoglutarate (alphaKG) dependent dioxygenase superfamily represent the largest class of non-heme iron oxidases and have important medical, ecological, and biotechnological roles. One such enzyme, taurine/alpha-ketoglutarate dioxygenase (TauD), catalyzes the conversion of 2-aminoethanesulfonat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0341096
更新日期:2003-05-20 00:00:00
abstract::High-resolution proton nuclear magnetic resonance (1H NMR) measurements were made on myosin, heavy meromyosin (HMM), myosin subfragment 1 (S1), light meromyosin (LMM), and actin. A strong signal from amino acid side chains undergoing motions too fast to be accounted for by simple rotations of groups on a rigid backbon...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00586a031
更新日期:1979-09-18 00:00:00
abstract::Structure-function studies of antibody-antigen systems include the identification of amino acid residues in the antigen that interact with an antibody and elucidation of their individual contributions to binding affinity. We used fluorescence correlation spectroscopy (FCS) and alanine-scanning mutagenesis to character...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0607047
更新日期:2006-11-28 00:00:00
abstract::In the course of measuring the concentrations of retinoic acids (RA) in bovine plasma, a major peak was observed which comigrated with 9-cis-RA on normal-phase high-performance liquid chromatography. Rechromatography of this retinoic on reverse-phase high-performance liquid chromatography showed that it was distinct f...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00004a013
更新日期:1995-01-31 00:00:00
abstract::Vitamin K dependent carboxylation of an exogenous peptide substrate and endogenous protein substrates, vitamin K epoxidation, and reduction of vitamin K epoxide were measured in subcellular fractions from rat liver. The rough microsomal fraction was highly enriched in all four activities; lower levels were found in sm...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00547a019
更新日期:1980-03-18 00:00:00
abstract::The bacterial DNA cytosine methyltransferase M.HhaI sequence-specifically modifies DNA in an S-adenosylmethionine dependent reaction. The enzyme stabilizes the target cytosine (GCGC) into an extrahelical position, with a concomitant large movement of an active site loop involving residues 80-99. We used multidimension...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi602662e
更新日期:2007-06-19 00:00:00
abstract::The interactions of mandelate racemase with divalent metal ion, substrate, and competitive inhibitors were investigated. The enzyme was found by electron paramagnetic resonance (EPR) to bind 0.9 Mn2+ ion per subunit with a dissociation constant of 8 muM, in agreement with its kinetically determined activator constant....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00677a006
更新日期:1975-03-25 00:00:00
abstract::The globular domain of chicken histone H1 (GH1) has been studied by 1H homonuclear and 1H-15N heteronuclear 2D NMR spectroscopy. After the full assignment of the proton and 15N resonances, the tertiary structure of GH1 was determined by an iterative procedure using distance geometry and restrained simulated annealing....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00203a004
更新日期:1994-09-20 00:00:00
abstract::The proton nuclear Overhauser effects of bovine alpha-lactalbumin were studied at 200 MHz by irradiation of an upfield ring current shifted methylene at -2.45 ppm (assigned to Ile-95) and two aromatic protons, Tyr-103 (8.36 ppm) and Trp-60 (5.85 ppm). The experimental results were consistent with a putative three-dime...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00346a035
更新日期:1985-12-03 00:00:00
abstract::In recent years, there has been dramatic growth in the study of RNA. RNA has gone from being known as an intermediate in the central dogma of molecular biology to a molecule with a large diversity of structure and function that is involved in all aspects of biology. As new functions are rapidly discovered, it has beco...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.7b01141
更新日期:2018-08-07 00:00:00
abstract::A 35-amino acid peptide corresponding to the putative "zinc finger" sequence of primase was prepared to study its zinc binding properties. When zinc was added to the peptide, it was found that the fluorescence quantum yield of the single tyrosine increased by 46% and the average lifetime by 34%. The binding stoichiome...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9612778
更新日期:1997-01-21 00:00:00
abstract::We have measured the hydration of CO2 catalyzed by bovine red cell carbonic anhydrase in mixtures of H2O and D2O at pH(D) which is in the plateau region of the pH--activity profile. The steady-state parameters for hydration Vmax and Km both show an exponential dependence on the atom fraction of deuterium in solvent wa...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00563a008
更新日期:1980-10-28 00:00:00
abstract::Azide binds to fast cytochrome bo with a stoichiometry of 1:1, the dissociation constant for this reaction being approximately 2 x 10(-5) M. The changes induced in the electronic absorption are very slight and are consistent with heme o remaining hexacoordinate high-spin, an observation confirmed by room temperature M...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961221d
更新日期:1996-10-29 00:00:00
abstract::The slowly hydrolyzed ATP analog adenosine 5'-(gamma-thiotriphosphate) (ATP gamma S) has been used in many studies of the muscle motor protein myosin in order to form a stable "weak binding" state analogous to the actin-S1-ATP complex However, the results from studies using ATP gamma S do not always agree with the res...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00049a018
更新日期:1995-12-12 00:00:00
abstract::R67 dihydrofolate reductase (DHFR) is an R-plasmid encoded enzyme that confers resistance to the antibacterial drug trimethoprim. This enzyme is not homologous in sequence or structure to chromosomal DHFRs. Equilibrium folding of tetrameric R67 DHFR was studied and found to be fully reversible. Formation of an inactiv...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00180a018
更新日期:1994-04-12 00:00:00
abstract::31P NMR spectra of equilibrium mixtures of enzyme-bound reactants and products of the adenylate kinase reaction (formula; see text) were analyzed by using computer simulations based on density matrix theory of chemical exchange. Since adenylate kinase has the unique feature that the reactants in the reverse direction ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00300a025
更新日期:1984-02-28 00:00:00
abstract::Translational initiation factor IF-2 is involved in a multistep pathway leading to the synthesis of the first peptide bond. IF-2 is a guanine nucleotide binding protein (G-protein) and catalyzes GTP hydrolysis in the presence of ribosomes. According to sequence homologies with other G-proteins, particularly EF-Tu, a t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00493a031
更新日期:1990-10-16 00:00:00
abstract::The effects of NH3 on the oxygen evolving enzyme have been investigated with EPR and steady-state O2 evolution. The following results were obtained. At low light intensity O2 evolution occurs in all centers even though ammonia is bound. This binding occurs in the S2 state and results in a modification of the multiline...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00453a003
更新日期:1990-01-09 00:00:00
abstract::The effect of Ca2+ binding on the circular dichroism (CD) and 270-MHz proton nuclear magnetic resonance (NMR) spectra of brain-specific S-100b calcium binding protein has been examined at two pH values, 8.5 and 7.5. At pH 8.5, S-100b protein binds two Ca2+ per monomer with Kd values of 6 x 10(-5) and 2 x 10(-4) M, whe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00276a033
更新日期:1983-03-29 00:00:00
abstract::We have used DNase I footprinting to examine the formation of antiparallel DNA triple helices on DNA fragments containing the homopurine target sites (GGA)2GGX(GGA)2GG.(CCT)2CCZ(CCT)2CC (where X.Z is each base pair in turn), with the GA- and GT-rich oligonucleotides, (GGA)2GGN(GGA)2GG and (GGT)2GGN(GGT)2GG (N = each b...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9609679
更新日期:1996-11-26 00:00:00
abstract::The importance of DNA polymerase-DNA minor groove interactions on translesion synthesis (TLS) was examined in vitro using variants of exonuclease-deficient Klenow polymerase and site-specifically modified DNA oligonucleotides. Polymerase variant R668A lacks primer strand interactions, while variant Q849A lacks templat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047591g
更新日期:2005-05-10 00:00:00
abstract::The spin-1/2-carrying NO molecule interacts with both the S1 and S2 states of the water oxidizing complex. The intermediates of the interaction can be resolved and trapped by NO treatment at subzero temperatures. At -30 degrees C and in the presence of approx. 500-700 microM NO, S1 loses the ability to yield by illumi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962899w
更新日期:1997-07-29 00:00:00