Abstract：
:Pantothenate synthetase (PS, EC 6.3.2.1) is the last enzyme in the pantothenate biosynthesis pathway, a metabolic pathway identified as a potential target for new antimicrobials. PS catalyzes the ATP-dependent condensation of pantoate and beta-alanine to form pantothenate. Here we report the overexpression, purification, enzyme assay, and tertiary structure of PS from Staphylococcus aureus. PS activity was experimentally confirmed, indicating a k(cat) value comparable to those of enzymes from other organisms. The structures of the apoenzyme and the reaction intermediate (pantoyl adenylate; PA) complex were determined by X-ray crystallography to resolutions of 2.5 and 1.85 A, respectively. Structural analysis indicated that the apoenzyme adopts an open and relatively mobile structure, while the complex structure is closed and entirely rigid. Structural comparison of the apoenzyme and the complex revealed how S. aureus PS undergoes open/close conformational change, and also determined the key interactions with the adenine ring of PA for a hinge bending domain closure. In the complex structure, PA and acetate are bound in the active site. We suggest that the acetate mimics the substrate beta-alanine. Therefore, the complex structure seems to represent a catalytic state poised for in-line nucleophilic attack on PA. These data also offer an alternative strategy for designing novel compounds that selectively inhibit PS activity.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Satoh A,Konishi S,Tamura H,Stickland HG,Whitney HM,Smith AG,Matsumura H,Inoue Tdoi
10.1021/bi1004206subject
Has Abstractpub_date
2010-08-03 00:00:00pages
6400-10issue
30eissn
0006-2960issn
1520-4995journal_volume
49pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:The glucose derivative, 2,2,6,6-tetramethylpiperidine-1-oxylglucose (TEMPO-glucose) was synthesized and examined for its ability to substitute for glucose as a substrate for the intestinal brush border membrane Na+/glucose cotransporter. TEMPO-glucose inhibited Na(+)-dependent phlorizin binding with an apparent KI of ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00231a012
更新日期：1991-04-30 00:00:00
abstract：:Bax protein plays a key role in mitochondrial membrane permeabilization and cytochrome c release upon apoptosis. Our recent data have indicated that the 20-residue C-terminal peptide of Bax (BaxC-KK; VTIFVAGVLTASLTIWKKMG), when expressed intracellularly, translocates to the mitochondria and exerts lethal effect on can...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi301195f
更新日期：2012-11-20 00:00:00
abstract：:The cAMP-dependent protein kinase (cAPK) plays an essential role during differentiation and fruit morphogenesis in Dictyostelium discoideum. The presence of an open reading frame on the gene, pkaC (previously named either Dd PK2 or Dd PK3 by different groups), predicts a 73-kDa polypeptide with 54% similarity to the c...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00088a003
更新日期：1993-09-21 00:00:00
abstract：:Recently, a new clathrin assembly protein (AP180) has been purified from coated vesicles of bovine brain (Ahle & Ungewickell, 1986). This protein has been shown to promote polymerization of clathrin into a homogeneous population of baskets under conditions where pure clathrin does not polymerize by itself. We have pur...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00416a040
更新日期：1988-08-09 00:00:00
abstract：:Arachidonate metabolites are potent biological mediators affecting multiple cellular functions. Although prostaglandins of the E series, which are products of the cyclooxygenase pathway, have been known as inhibitors or down-regulators of fibroblast proliferation and collagen synthesis, the more recently discovered pr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00408a028
更新日期：1988-04-19 00:00:00
abstract：:The physiological effects of anesthetics have been ascribed to their interaction with hydrophobic sites within functionally relevant CNS proteins. Studies have shown that volatile anesthetics compete for luciferin binding to the hydrophobic substrate binding site within firefly luciferase and inhibit its activity (Fra...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi800544d
更新日期：2008-09-09 00:00:00
abstract：:Two heat-shock proteins that show high identity with the Escherichia coli chaperonin 60 (groEL) and chaperonin 10 (groES) chaperonin proteins were purified and characterized from photolithoautotrophically grown Rhodobacter sphaeroides. The proteins were purified by using sucrose density gradient centrifugation and Mon...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00247a013
更新日期：1991-08-20 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：:The location of the sites of methylation of the methyl-accepting chemotaxis proteins (MCP) of Escherichia coli has been investigated by trypsin digestion and mapping of the peptides. The two principal MCPs, MCP I and MCP II, were found to have very similar methyl-labeled peptide patterns. Each produced three major spe...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00507a051
更新日期：1981-02-17 00:00:00
abstract：:Light-harvesting complex II (LHC-II) regulates the light energy distribution between photosystem I and II in plants. This process is mediated by phosphorylation of the LHC-II protein, which depends on the oxidation state of photosynthetic electron carriers. In addition to this regulatory mechanism, it has recently bee...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi034114+
更新日期：2003-09-02 00:00:00
abstract：:The complete coordination scheme for Mn(II) in transition-state-analogue complexes with creatine kinase has been determined by electron paramagnetic resonance (EPR) spectroscopy. Perturbations in the EPR spectra for Mn(II) due to superhyperfine coupling to 17O of selectively labeled ligands have been used to identify ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00565a002
更新日期：1980-11-25 00:00:00
abstract：:The solution structure of the self-complementary dodecamer 5'd(CGCGPATTCGCG)2, containing a purine-thymine base pair within the hexameric canonical recognition site GAATTC for the restriction endonuclease EcoRI, is investigated by nuclear magnetic resonance spectroscopy and restrained molecular dynamics. Nonexchangeab...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00411a042
更新日期：1988-05-31 00:00:00
abstract：:The one-for-one exchange of Cl(-) and HCO(3)(-) ions is catalyzed by human erythrocyte anion exchanger 1 (hAE1) through a ping-pong mechanism whereby the protein exists in two main conformations, with the single anion-binding site exposed at either the cytoplasmic (inner) side (E(i)) or the extracellular side (E(o)), ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051916r
更新日期：2006-05-23 00:00:00
abstract：:A substrate analogue, 6-(difluoromethyl)tryptophan, was developed and characterized for mechanistic investigation of tryptophanase. The utility of this derivative was based on its ability to partition between fluoride elimination and carbon-carbon bond scission during tryptophan metabolism. The non-enzymatic hydrolysi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00221a018
更新日期：1991-02-19 00:00:00
abstract：:The interaction of 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase, 4,5-dideoxyshikimate 3-phosphate (ddS3P), and [2-13C]-and [3-13C]phosphoenolpyruvate (PEP) has been examined by 13C NMR spectroscopy. Although no resonances due to a dead-end intermediate complex could be detected, an enzyme active site specific fo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00446a003
更新日期：1989-10-03 00:00:00
abstract：:The sucrose (CscB) permease belongs to the oligosaccharide:H(+) symporter family of the Major Facilitator Superfamily and is homologous to the lactose permease from Escherichia coli. Sucrose transport in cells expressing sucrose permease is completely inhibited by N-ethylmaleimide (NEM), suggesting that one or more of...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi000124o
更新日期：2000-05-23 00:00:00
abstract：:The hydrophobic, membrane-binding domain of purified human erythrocyte acetylcholinesterase was labeled with the photoactivated reagent 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine. The radiolabel was incorporated when the enzyme was prepared in detergent-free aggregates, in detergent micelles, or in phospholip...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00359a004
更新日期：1986-06-03 00:00:00
abstract：:A simple and rapid procedure is described for purification of P-glycoprotein (Pgp) from a multidrug-resistant Chinese hamster ovary cell line (CR1R12) in which the plasma membranes are highly enriched in Pgp (Al-Shawi, M.K., Senior A.E. (1993) J. Biol. Chem, 268, 4197-4206). The procedure consisted of octylglucoside s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00189a008
更新日期：1994-06-14 00:00:00
abstract：:The insulin hexamer is an allosteric protein widely used in formulations for the treatment of diabetes. The hexamer exhibits positive and negative cooperativity and apparent half-site binding activity, reflecting the interconversion of three allosteric states, designated as T6, T3R3, and R6. The hexamer contains two s...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi050392s
更新日期：2005-05-31 00:00:00
abstract：:The head of the P22 bacteriophage is interrupted by a unique dodecameric portal vertex that serves as a conduit for the entrance and exit of the DNA. Here, the in vitro unfolding/refolding processes of the portal protein of P22 were investigated at different temperatures (1, 25, and 37 degrees C) through the use of ur...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi700006d
更新日期：2007-06-26 00:00:00
abstract：:The locations of RNA-RNA cross-links in 16S rRNA were determined after in vivo incorporation of 4-thiouridine (s(4)U) into RNA in a strain of Escherichia coli deficient in pyrimidine synthesis and irradiation at >320 nm. This was done as an effort to find RNA cross-links different from UVB-induced cross-links that wou...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi049702h
更新日期：2004-07-20 00:00:00
abstract：:Although the membrane-associated surfactant protein B (SP-B) is an essential component of lung surfactant, which is itself essential for life, the molecular basis for its activity is not understood. SP-B's biophysical functions can be partially mimicked by subfragments of the protein, including the C-terminus. We have...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0481895
更新日期：2004-12-07 00:00:00
abstract：:The nonspecific phospholipase C from Bacillus cereus is a zinc metalloenzyme that catalyzes the hydrolysis of phospholipids to yield diacylglycerol and a phosphate monoester. Glu-4 has been proposed as a potential candidate for the general base in the hydrolysis reaction and was shown to interact with the substrate he...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi972751s
更新日期：1998-03-24 00:00:00
abstract：:Trytophanase from Bacillus alvei was resolved from its coenzyme, pyridoxal phosphate, by treatment with cysteine followed by column chromatography. Spectrophotometric titration of apoenzyme with pyridoxal-P showed 1 mol of pyridoxal-P bound per 52,000 g of enzyme. Kinetic analysis of coenzyme binding showed hyperbolic...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00690a024
更新日期：1975-09-23 00:00:00
abstract：:Phoborhodopsin from Halobacterium salinarum (salinarum phoborhodopsin, spR also called HsSR II) is a photoreceptor for the negative phototaxis of the bacterium. A unique feature of spR is the formation of a shorter wavelength photoproduct, P480, observed at liquid nitrogen temperature beside the K intermediate. Format...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi200598r
更新日期：2011-08-23 00:00:00
abstract：:The intestinal fatty acid binding protein (IFABP) is a small (15 kDa) protein consisting mostly of 10 antiparallel beta-strands (A-J) and a small helical region that serves as a portal for the ligand. Two beta-sheet structures (strands A-E and F-J) surround a cavity into which the ligand binds. In this work, we invest...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0301688
更新日期：2003-10-28 00:00:00
abstract：:X-ray crystallographic data to 1.9-A resolution were collected on sperm whale metmyoglobin equilibrated with 7 atm of xenon gas. The results indicate four xenon sites of occupancy from 0.45 to 1.0. These sites are located in interior spaces or packing defects of the myoglobin molecule. The effects of the bound xenon o...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00308a002
更新日期：1984-06-19 00:00:00
abstract：:Sulfhydryl-reactive derivatives of glucosamine were synthesized as potentially transportable affinity labels of the human erythrocyte hexose carrier. N-Maleoylglycyl derivatives of either 6- or 2-amino-2-deoxy-D-glucopyranose were the most potent inhibitors of 3-O-methylglucose uptake, with concentrations of half-maxi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00430a044
更新日期：1989-02-21 00:00:00
abstract：:Because oxygen and selenium are in the same group (Family VI) in the periodic table, the site-specific mutagenesis at the atomic level by replacing RNA oxygen with selenium can provide insights on the structure and function of catalytic RNAs. We report here the first Se-derivatized ribozymes transcribed with all nucle...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi060455m
更新日期：2006-07-25 00:00:00
abstract：:Trp151 in the lactose permease of Escherichia coli (LacY) is an important component of the sugar-binding site and the only Trp residue out of six that is in close proximity to the galactopyranoside in the structure (1PV7). The short distance between Trp151 and the sugar is favorable for Förster resonance energy transf...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi061632m
更新日期：2006-12-26 00:00:00
