Abstract:
:The enzyme RhlI catalyzes the formation of N-butyrylhomoserine lactone from S-adenosylmethionine and N-butyrylacyl carrier protein. N-Butyrylhomoserine lactone serves as a quorum-sensing signal molecule in Pseudomonas aeruginosa, and is implicated in the regulation of many processes involved in bacterial virulence and infectivity. The P. aeruginosa genome contains three genes encoding acyl carrier proteins. We have cloned all three genes, expressed the acyl carrier proteins, and characterized each as a substrate for RhlI. A continuous, spectrophotometric assay was developed to facilitate kinetic and mechanistic studies of RhlI. Acp1, which has not been characterized previously, was a good substrate for RhlI, with a K(m) of 7 microM; the reaction proceeded with a k(cat) value of 0.35 s(-1). AcpP, which supports fatty acid biosynthesis, was also a good substrate in the RhlI reaction, where k(cat) was 0.46 s(-1), and the K(m) for AcpP was 6 microM. The third acyl carrier protein, Acp3, was a poor substrate for RhlI, with a K(m) of 280 microM; k(cat) was 0.03 s(-1). Taken together with microarray data from the literature which show that expression of the gene encoding Acp1 is under the control of the quorum-sensing system, our data suggest that Acp1 is likely to be the substrate for RhlI in vivo. Isotope labeling studies were conducted to investigate the chemical mechanism of the RhlI-catalyzed lactonization reaction. Solvent deuterons were not incorporated into product, which implicates a direct attack mechanism in which the carboxylate oxygen of the presumptive N-butyryl-SAM intermediate attacks the methylene carbon adjacent to the sulfonium ion. Alternative mechanisms, in which N-butyrylvinylglycine is formed via elimination of methylthioadenosine, were ruled out on the basis of the observation that RhlI failed to convert authentic N-butyrylvinylglycine to N-butyryl-L-homoserine lactone.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Raychaudhuri A,Jerga A,Tipton PAdoi
10.1021/bi048005msubject
Has Abstractpub_date
2005-03-01 00:00:00pages
2974-81issue
8eissn
0006-2960issn
1520-4995journal_volume
44pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::To investigate the effect of a reduced level of selenocysteine (Sec) tRNA[Ser]Sec in selenoprotein biosynthesis, two mouse embryonic stem (ES) cell lines heterozygous for the corresponding gene were generated by homologous recombination of the host genome with targeting vectors encoding a deleted or a disrupted tRNA[S...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970608t
更新日期:1997-07-15 00:00:00
abstract::Prion diseases are associated with a conformational switch in the prion protein (PrP) from its normal cellular form (denoted PrP(C)) to a disease-associated "scrapie" form (PrP(Sc)). A number of PrP(Sc)-like conformations can be generated by incubating recombinant PrP(C) at low pH, indicating that protonation of key r...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100572j
更新日期:2010-10-12 00:00:00
abstract::The fungal neurotoxin alpha-cyclopiazonic acid (CPA), a nanomolar inhibitor of Ca(2+)-ATPase with a unique pentacyclic indole tetramic acid scaffold, is assembled by a three enzyme pathway CpaS, CpaD, and CpaO in Aspergillus sp. We recently characterized the first pathway-specific enzyme CpaS, a hybrid two module poly...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901597j
更新日期:2009-11-24 00:00:00
abstract::The role of pyridoxal 5'-phosphate (PLP) in the radical-mediated amino group migration catalyzed by lysine 2,3-aminomutase from Clostridia SB4 has been investigated by electron spin echo envelope modulation (ESEEM) spectroscopy. This pulsed electron paramagnetic resonance (EPR) method was used to estimate the distance...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00031a033
更新日期:1995-08-08 00:00:00
abstract::Protein kinase C and two other proteins with molecular masses of 64 and 32 kDa, purified from bovine brain, constitute a type of protein that binds a large number of calcium ions in a phospholipid-dependent manner. This study suggested that these proteins also induced extensive clustering of acidic phospholipids in th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00246a013
更新日期:1991-08-13 00:00:00
abstract::The sensitivity of the fluorescence energy of 2-p-toluidinylnaphthalene-6-sulfonate (TNS) to apparent solvent polarity is considerably greater in methanol-water mixtures and lower primary alcohols than in higher primary alcohols or solvents containing no hydroxyl groups. It is suggested that the behavior of TNS in low...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00675a016
更新日期:1975-02-25 00:00:00
abstract::To identify regions of the Torpedo Na,K-ATPase alpha-subunit that interact with membrane lipid and to characterize conformationally dependent structural changes in the transmembrane domain, we have proteolytically mapped the sites of photoincorporation of the hydrophobic compounds 3-(trifluoromethyl)-3-(m-[(125)I]iodo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0015461
更新日期:2000-11-07 00:00:00
abstract::Anthrax toxin receptor 1 (ANTXR1)/tumor endothelial marker 8 (TEM8) is one of two known proteinaceous cell surface anthrax toxin receptors. A metal ion dependent adhesion site (MIDAS) present in the integrin-like inserted (I) domain of ANTXR1 mediates the binding of the anthrax toxin subunit, protective antigen (PA). ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100887w
更新日期:2010-08-31 00:00:00
abstract::Little is known about the extent to which protein flexibility contributes to antigen-antibody recognition and cross-reactivity. Using short coil peptides (leucine zippers) as model antigens, we demonstrate that a monoclonal antibody can force a noncognate peptide into a conformation that is similar to the conformation...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00050a035
更新日期:1995-12-19 00:00:00
abstract::A modified form of procollagen was extracted with 10 M urea from the skin of lambs with dermatosparaxis, a disease which is produced by a genetic defect in the conversion of procollagen to collagen. The extracts contained little if any alpha1 and alpha2 chains of normal type I collagen, and instead they contained the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00658a024
更新日期:1976-06-29 00:00:00
abstract::ncd is a kinesin-related motor protein from Drosophila that moves in the opposite direction along microtubules to kinesin. To learn more about the ncd mechanism, ncd motor domain (R335-K700) was expressed in Escherichia coli and its enzymatic characteristics were studied. The ncd motor domain was purified from the cel...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00040a042
更新日期:1995-10-10 00:00:00
abstract::Isolated alpha and beta chains from Xenopus laevis hemoglobin have been purified. The isolation procedure yields native alpha chains whose functional behavior has been characterized and compared with that of human alpha chains. Isolated beta chains in the presence of oxygen are characterized by low stability, and henc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00395a022
更新日期:1987-10-20 00:00:00
abstract::The newly described adapter molecule p130 Crk-associated substrate (Cas) has been reported to contribute to cytoskeletal organization through assembly of actin filaments and to be pivotal in embryonic development and in oncogene-mediated transformation. We characterized the regulation of Cas tyrosine phosphorylation i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi991849z
更新日期:2000-05-16 00:00:00
abstract::Trypsin and N-[3-[4-[4-(amidinophenoxy)carbonyl]phenyl]-2-methyl-2-propenoyl]- N-allylglycine methanesulfonate (1), a newly designed and orally active synthetic trypsin inhibitor, were cocrystallized. The space group of the crystal is P2(1)2(1)2(1) with cell constants a = 63.74 A, b = 63.08 A, and c = 69.38 A, which i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00039a046
更新日期:1995-10-03 00:00:00
abstract::Actin-depolymerizing factor (ADF)/cofilin enhances the turnover of actin filaments by two separable activities: filament severing and pointed-end depolymerization. Multicellular organisms express multiple ADF/cofilin isoforms in a tissue-specific manner, and the vertebrate proteins are grouped into ADFs and cofilins o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050933d
更新日期:2005-11-01 00:00:00
abstract::Urease from Klebsiella aerogenes [Jabri et al. (1995) Science 268, 998-1004] is an (alpha beta gamma)3 trimer with each alpha-subunit having an (alpha beta)8-barrel domain containing a binickel active center. Here we examine structure-function relations for urease in more detail through structural analysis of the urea...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960424z
更新日期:1996-08-20 00:00:00
abstract::F(1)-ATPase mutations in Escherichia coli that changed the strength of hydrogen bonds between the alpha and beta subunits in a location that links the catalytic site to the interface between the beta catch loop and the gamma subunit were examined. Loss of the ability to form the hydrogen bonds involving alphaS337, bet...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi052592+
更新日期:2006-09-19 00:00:00
abstract::We assigned l-talarate dehydratase (TalrD) and galactarate dehydratase (GalrD) functions to a group of orthologous proteins in the mechanistically diverse enolase superfamily, focusing our characterization on the protein encoded by the Salmonella typhimurium LT2 genome (GI:16766982; STM3697). Like the homologous mande...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7008882
更新日期:2007-08-21 00:00:00
abstract::The salivary antimicrobial peptide histatin-5 is able to aggregate and fuse negatively charged small unilamellar vesicles, and this fusogenic activity is selectively induced by the presence of zinc ions. Circular dichroism spectroscopy shows that histatin-5, in the presence of negatively charged vesicles and zinc ions...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990212c
更新日期:1999-07-27 00:00:00
abstract::Retinitis pigmentosa (RP), a neurodegenerative disorder, can arise from single point mutations in rhodopsin, leading to a cascade of protein instability, misfolding, aggregation, rod cell death, retinal degeneration, and ultimately blindness. Divalent cations, such as zinc and copper, have allosteric effects on misfol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi800968w
更新日期:2009-03-03 00:00:00
abstract::From a series of 15N-resolved 3D ROESY-HMQC and 13C-resolved 3D NOESY-HMQC spectra of the FK506 binding protein (FKBP)/ascomycin complex in H2O, the locations of three tightly bound water molecules were identified. These waters are all buried within the interior of the complex and form an integral part of its structur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00061a004
更新日期:1993-03-16 00:00:00
abstract::4-Chlorobenzoyl-coenzyme A (4-CBA-CoA) dehalogenase catalyzes the hydrolysis of 4-CBA-CoA to 4-hydroxybenzoyl-coenzyme A (4-HBA-CoA) via a nucleophilic aromatic substitution pathway involving the participation of an active site carboxylate side chain in covalent catalysis. In this paper we report on the identification...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9609533
更新日期:1996-08-20 00:00:00
abstract::Laser light scattering has been applied to a systematic study of a heterogeneous solution of tubulin at low temperature--conditions under which tubulin assembly into microtubules does not take place. Methods of analyzing laser light scattering results obtained from solutions containing multiple components are discusse...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00645a020
更新日期:1977-12-27 00:00:00
abstract::Modified (Arg63-Ile64 reactive-site peptide bond hydrolyzed) soybean trypsin inhibitor (Kunitz) with all reactive amino groups, except that of Ile64, protected was described in the preceding paper (Kowalski, D., and Laskowski, M., Jr. (1976), Biochemistry, preceding paper in this issue). Treatment of this inhibitor wi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00651a022
更新日期:1976-03-23 00:00:00
abstract::The role of interleukin-8 (IL-8) and related CXC chemokines has been demonstrated in many human diseases. However, more profound studies, e.g., by blocking the effect of these inflammatory mediators, request animal models and hence the identification of all human counterparts for commonly used laboratory animals. In t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0011227
更新日期:2000-11-28 00:00:00
abstract::The enzyme As(III) S-adenosylmethionine methyltransferase (EC 2.1.1.137) (ArsM or AS3MT) is found in members of every kingdom, from bacteria to humans. In these enzymes, there are three conserved cysteine residues at positions 72, 174, and 224 in the CmArsM orthologue from the thermophilic eukaryotic alga Cyanidioschy...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201500c
更新日期:2012-02-07 00:00:00
abstract::Chemical and enzymatic probing (footprinting) of the reactivity of the promoter DNA backbone is applied to characterize two binary open complexes RPo1 (-Mg2+) and RPo2 (+Mg2+), formed by Escherichia coli RNA polymerase (E sigma 70) at the lambda PR promoter. We report that HO. detects major differences in backbone rea...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00048a004
更新日期:1995-12-05 00:00:00
abstract::The monoamine transporter of the chromaffin granule membranes can be specifically labeled by the photoaffinity reagent 7-azido-8-[125I]iodoketanserin. The characteristics of the labeled protein have been investigated. Two-dimensional gel electrophoresis of the labeled membranes indicated a MW of about 70,000 and an is...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00122a012
更新日期:1992-02-25 00:00:00
abstract::Despite the expression of only one type of receptor, there is great variation in the ability of different cell types to discriminate between C5a and its more stable metabolite, C5a des Arg74. The mechanism that underlies this phenomenon is not understood but presumably involves differences in the interaction with the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990139q
更新日期:1999-07-27 00:00:00
abstract::cGMP is the natural activator of the cyclic nucleotide-gated channel originally isolated from rod photoreceptors but now known to be expressed in a wide variety of neural and non-neural cells. To identify antagonists of cGMP action and to better understand the interaction between cGMP and the channel protein, experime...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961763v
更新日期:1996-12-24 00:00:00