Abstract:
:The substrate specificity of the beta-glucosidase (CelB) from the hyperthermophilic archaeon Pyrococcus furiosus, a family 1 glycosyl hydrolase, has been studied at a molecular level. Following crystallization and X-ray diffraction of this enzyme, a 3.3 A resolution structural model has been obtained by molecular replacement. CelB shows a homo-tetramer configuration, with subunits having a typical (betaalpha)(8)-barrel fold. Its active site has been compared to the one of the previously determined 6-phospho-beta-glycosidase (LacG) from the mesophilic bacterium Lactococcus lactis. The overall design of the substrate binding pocket is very well conserved, with the exception of three residues that have been identified as a phosphate binding site in LacG. To verify the structural model and alter its substrate specificity, these three residues have been introduced at the corresponding positions in CelB (E417S, M424K, F426Y) in different combinations: single, double, and triple mutants. Characterization of the purified mutant CelB enzyme revealed that F426Y resulted in an increased affinity for galactosides, whereas M424K gave rise to a shifted pH optimum (from 5.0 to 6.0). Analysis of E417S revealed a 5-fold and a 3-fold increase of the efficiency of hydrolyzing o-nitrophenol-beta-D-galactopyranoside-6-phosphate, in the single and triple mutants, respectively. In contrast, their activity on nonphosphorylated sugars was largely reduced (30-300-fold). The residue at position E417 in CelB seems to be the determining factor for the difference in substrate specificity between the two types of family 1 glycosidases.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Kaper T,Lebbink JH,Pouwels J,Kopp J,Schulz GE,van der Oost J,de Vos WMdoi
10.1021/bi992463rsubject
Has Abstractpub_date
2000-05-02 00:00:00pages
4963-70issue
17eissn
0006-2960issn
1520-4995pii
bi992463rjournal_volume
39pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The molecular weight of phosphophoryn, an acidic phosphoprotein unique to dentin matrix, has been difficult to determine because of a combination of neutral protease activities in this tissue and the intrinsic high charge density of the molecule. In this study, bovine dentin phosphophoryn (BDPP) was isolated by a proc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00287a025
更新日期:1983-08-30 00:00:00
abstract::The flavocytochrome p-cresol methylhydroxylase from Pseudomonas putida has been reported to have a Mr of 114,000 and to consist of two subunits, a flavoprotein and a cytochrome c, each with a Mr of 58,000. Recent X-ray crystallographic data from our laboratories [Shamala, N., Lim, L. W., Mathews, F. S., McIntire, W., ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00368a021
更新日期:1986-10-07 00:00:00
abstract::Few techniques can identify interactions between proteins and individual water molecules when the protein is in solution. The present work has sought to bridge the gap between the molecular level studies and the search for a physical property of the solution (bathing the proteins) that would regulate the protein hydra...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi012210+
更新日期:2002-08-13 00:00:00
abstract::Because of its ability to selectively glucosylate misfolded glycoproteins, UDP-glucose:glycoprotein glucosyltransferase (UGGT) functions as a folding sensor in the glycoprotein quality control system in the endoplasmic reticulum (ER). The unique property of UGGT derives from its ability to transfer a glucose residue t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00785
更新日期:2015-08-11 00:00:00
abstract::The P-glycoprotein multidrug transporter functions as an ATP-driven efflux pump for a large number of structurally unrelated hydrophobic compounds. Substrates are believed to gain access to the transporter after partitioning into the membrane, rather than from the extracellular aqueous phase. The binding of drug subst...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990064q
更新日期:1999-05-25 00:00:00
abstract::A survey of RuvB protein-mediated ATP hydrolysis yields the following observations. (1) The RuvB protein exhibits a DNA-independent ATPase activity with a turnover number (based on a RuvB monomer) approaching 6 min-1 and a Km of 154 microM. Single-stranded DNA and linear duplex DNA have small but significant effects o...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00030a019
更新日期:1995-08-01 00:00:00
abstract::Studies of the mechanism of protein secretion in a Gram-positive bacterium, Bacillus subtilis, yielded results very similar to those previously obtained with a Gram-negative organism: nascent chains protruding from protoplasts could be labeled extracellularly; the labeled chains could be recovered on polysomes isolate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00568a030
更新日期:1979-01-09 00:00:00
abstract::Evidence is presented that indicates tropoelastin is derived from a soluble elastin with a molecular weight of 95000. Tropoelastin and its proposed precursor were isolated from the aortas of copper-deficient chicks. Although it is doubtful that the proposed precursor is an initial product of elastin translation, i.e.,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00585a004
更新日期:1979-09-04 00:00:00
abstract::Reactivation of organophosphate (OP)-inhibited acetylcholinesterase (AChE) is a key objective in the treatment of OP poisoning. This study with native, wild-type, and mutant recombinant DNA-expressed AChEs, each inhibited by representative OP compounds, establishes a relationship between edrophonium acceleration of ox...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9905720
更新日期:1999-08-03 00:00:00
abstract::Methylmalonyl-CoA mutase from Propionibacterium shermanii is an adenosylcobalamin-dependent enzyme which catalyzes the reversible isomerization of methylmalonyl-CoA and succinyl-CoA. The rate of tritium loss from 5'-[3H]adenosylcobalamin during the enzymic reaction and the relative rates of tritium appearance in subst...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961250o
更新日期:1996-09-10 00:00:00
abstract::To gain insight into the folding of large proteins, we constructed a bovine beta-lactoglobulin (beta-lg) dimeric mutant, A34C/C121A beta-lg. In the mutant, a free thiol group of wild-type beta-lg at Cys121 was removed and two beta-lg molecules were linked by a disulfide bridge through Cys34 created at the dimer's inte...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi8001715
更新日期:2008-06-03 00:00:00
abstract::Calcium and its receptor protein calmodulin function in the regulation of proliferation of mammalian cells. A 68 kDa calmodulin-specific binding protein was shown previously to be associated with growth factor-dependent progression of a variety of mammalian cells from G1 to S phase and to stimulate DNA synthesis in pe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00012a002
更新日期:1995-03-28 00:00:00
abstract::Binding of streptokinase (SK) to plasminogen (Pg) activates the zymogen conformationally and initiates its conversion into the fibrinolytic proteinase, plasmin (Pm). Equilibrium binding studies of SK interactions with a homologous series of catalytic site-labeled fluorescent Pg and Pm analogues were performed to resol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000594i
更新日期:2000-11-14 00:00:00
abstract::Horseradish peroxidase (HRP) was oxidized by IrCl6(2-) to a mixture of compounds I and II, the rate of oxidation and the ratio of the mixture being greatly affected by pH (Hayashi & Yamazaki, 1979). Oxidation of HRP by IrCl6(2-) in the presence of fluoride was significantly accelerated. This resulted in the formation ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00046a003
更新日期:1995-11-21 00:00:00
abstract::Quadruplex-specific molecules can serve as suitable drugs in cancer therapy. We have synthesized a pair of furan-based cyclic homooligopeptides, ligand 1 and ligand 2, to specifically target G-quadruplexes. We have shown by CD spectroscopy and UV melting that these ligands can effectively induce G-quadruplex structure...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005927
更新日期:2010-09-28 00:00:00
abstract::Chicken liver phosphoenolpyruvate carboxykinase (PEPCK) is activated by Cr2+ as the sole activator under anaerobic conditions. PEPCK was modified with Cr3+, starting with either Cr2+ or Cr3+. Cr3+ has the distinct advantage of being a paramagnetic cation that could serve as a paramagnetic probe. Activators Mn2+, Mg2+,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9807299
更新日期:1998-06-02 00:00:00
abstract::The structure of a synthetic peptide corresponding to residues 1-45 of rabbit muscle adenylate kinase has been studied in aqueous solution by two-dimensional NMR, FTIR, and CD spectroscopy. This peptide, which binds MgATP and is believed to represent most of the MgATP-binding site of the enzyme [Fry, D.C., Kuby, S.A.,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00410a009
更新日期:1988-05-17 00:00:00
abstract::4-Oxalocrotonate tautomerase (4-OT) and YwhB, a 4-OT homologue found in Bacillus subtilis, exhibit a low level hydratase activity that converts trans-3-haloacrylates to acetaldehyde, presumably through a malonate semialdehyde intermediate. The mechanism for the initial transformation of the 3-haloacrylate to malonate ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035495u
更新日期:2004-01-27 00:00:00
abstract::Lytic polysaccharide monooxygenases (LPMOs) are ubiquitous oxidoreductases, facilitating the degradation of polymeric carbohydrates in biomass. Cellobiose dehydrogenase (CDH) is a biologically relevant electron donor in this process, with the electrons resulting from cellobiose oxidation being shuttled from the CDH de...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b01178
更新日期:2019-03-05 00:00:00
abstract::The solution conformation of an agalactosyl penta-antennary "bisected" N-linked glycan from hen ovomucoid has been determined using a combination of 1H-NMR NOE measurements and restrained molecular dynamics (MD) simulations. The majority of glycosidic linkages exhibited restricted torsional fluctuations about the glob...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00043a018
更新日期:1995-10-31 00:00:00
abstract::Activated receptors with a tyrosine kinase activity induce a variety of responses like changes in the differentiation and mitogenic status of cells. These responses are mediated in part by p21ras. Some of these activated receptors induce in certain cell types a pronounced, but transient, increase in Ras-GTP. We have s...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00189a053
更新日期:1994-06-14 00:00:00
abstract::A chromatin acceptor protein for the avian oviduct progesterone receptor (PR), termed receptor binding factor 1 (RBF-1), has recently been shown to (1) be a component of the nuclear binding sites (acceptor sites) for PR and (2) generate high-affinity binding sites (termed the RBF-1 class of sites) on avian genomic DNA...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00103a019
更新日期:1991-10-01 00:00:00
abstract::Human heart short chain L-3-hydroxyacyl-CoA dehydrogenase (SCHAD) catalyzes the oxidation of the hydroxyl group of L-3-hydroxyacyl-CoA to a keto group, concomitant with the reduction of NAD+ to NADH, as part of the beta-oxidation pathway. The homodimeric enzyme has been overexpressed in Escherichia coli, purified to h...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9829027
更新日期:1999-05-04 00:00:00
abstract::Prodan [6-propionyl-2-(dimethylamine)naphthalene] is a hydrophobic fluorescent probe which is extremely sensitive to both the polarity and the hydrogen-bond donating capacity of the solvent. In binary mixtures of solvents, the hydrogen-bond donating effect on Prodan fluorescence saturates at relatively low concentrati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00154a021
更新日期:1992-10-06 00:00:00
abstract::Isotope exchange studies show that beef liver fructokinase has a random kinetic mechanism in which release of fructose from the enzyme is slower than that catalytic reaction. The stickiness of fructose in the presence of MgATP is confirmed by isotope partition studies, which show it to be released 0.53 times as fast a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00629a021
更新日期:1977-05-17 00:00:00
abstract::The equilibrium between the rat brain tubulin alpha beta dimer and the dissociated alpha and beta monomers has been studied by analytical ultracentrifugation with use of a new method employing short solution columns, allowing rapid equilibration and hence short runs, minimizing tubulin decay. Simultaneous analysis of ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00228a023
更新日期:1991-04-09 00:00:00
abstract::Pyruvate phosphate dikinase catalyzes the interconversion of adenosine 5'-triphosphate (ATP), orthophosphate (P(i)), and pyruvate with adenosine 5'-monophosphate (AMP), pyrophosphate (PP(i)), and phosphoenolpyruvate (PEP). The Arg 561 residue of Clostridium symbiosum PPDK is contained within a Gly-rich stretch of sequ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00007a012
更新日期:1995-02-21 00:00:00
abstract::The synthesis and secretion of several acute-phase proteins increases markedly following physiological stress. alpha 1-Acid glycoprotein (AGP), a major acute-phase reactant made by the liver, is strongly induced by inflammatory agents such as lipopolysaccharide (LPS). Nuclear run-on assay showed a 17-fold increase in ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00122a005
更新日期:1992-02-25 00:00:00
abstract::Bacterial resistance to the aminoglycoside antibiotics is manifested primarily through the production of enzymes which covalently modify these drugs. The Enterococci and Staphylococci produce an ATP-dependent kinase, APH(3')-IIIa, which phosphorylates such antibiotics as kanamycin, amikacin, and neomycin, and this enz...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9603884
更新日期:1996-07-02 00:00:00
abstract::Human lecithin:cholesterol acyltransferase (LCAT, E.C.2.3.1.43) is a serine-type esterase that contains six cysteines, two of which, Cys31 and Cys184, are free. The remaining cysteines form disulfide links. One of these is between Cys50 and Cys74 and the other is between Cys313 and Cys356. The cDNA of LCAT and mutants...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00063a021
更新日期:1993-03-30 00:00:00