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 transfer (FRET) to nitrophenyl or dansyl derivatives with the fluorophore at the anomeric position of galactose. Modeling of 4-nitrophenyl-alpha-d-galactopyranoside (alpha-NPG) in the binding-site of LacY places the nitrophenyl moiety about 12 A away from Trp151, a distance commensurate with the Förster distance for a Trp-nitrobenzoyl pair. We demonstrate here that alpha-NPG binding to LacY containing all six native Trp residues causes galactopyranoside-specific FRET from Trp151. Moreover, binding of alpha-NPG is sufficiently slow to resolve time-dependent fluorescence changes by stopped-flow. The rate of change in Trp --> alpha-NPG FRET is linearly dependent upon sugar concentration, which allows estimation of kinetic parameters for binding. Furthermore, 2-(4'-maleimidylanilino)naphthalene-6-sulfonic acid (MIANS) covalently attached to the cytoplasmic end of helix X is sensitive to sugar binding, reflecting a ligand-induced conformational change. Stopped-flow kinetics of Trp --> alpha-NPG FRET and sugar-induced changes in MIANS fluorescence in the same protein reveal a two-step process: a relatively rapid binding step detected by Trp151 --> alpha-NPG FRET followed by a slower conformational change detected by a change in MIANS fluorescence.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Smirnova IN,Kasho VN,Kaback HRdoi
10.1021/bi061632msubject
Has Abstractpub_date
2006-12-26 00:00:00pages
15279-87issue
51eissn
0006-2960issn
1520-4995journal_volume
45pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The nuclear transport of the chromatin remodeling protein nucleoplasmin and chromatin building histones is mediated by importins. Nucleoplasmin (NP) contains a classical bipartite nuclear localization signal (NLS) that is recognized by the importin α/β heterodimer, while histones present multiple NLS-like motifs that ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2008867
更新日期:2011-08-23 00:00:00
abstract::The gene coding for the enzyme 2,3-dihydroxybenzoate-AMP ligase (2,3DHB-AMP ligase), responsible for activating 2,3-dihydroxybenzoic acid in the biosynthesis of the siderophore enterobactin, has been subcloned into the multicopy plasmid pKK223-3 and overproduced in a strain of Escherichia coli. The protein is an alpha...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00443a008
更新日期:1989-08-22 00:00:00
abstract::The clade B/intracellular serpins protect cells from peptidase-mediated injury by forming covalent complexes with their targets. SERPINB12 is expressed in most tissues, especially at cellular interfaces with the external environment. This wide tissue distribution pattern is similar to that of granzyme A (GZMA). Becaus...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b01042
更新日期:2015-11-17 00:00:00
abstract::Lactose repressor protein, regulator of lac enzyme expression in Escherichia coli, maintains its structure and function at extremely low protein concentrations (<10(-)12 M). To examine the unfolding and dissociation of this tetrameric protein, structural transitions in the presence of varying concentrations of urea we...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9900727
更新日期:1999-05-18 00:00:00
abstract::The kinetics of ATP-induced Ca2+ uptake by vesicular dispersions of sarcoplasmic reticulum were determined with a time resolution of about 10 ms, depending on the temperature. Ca2+ uptake was initiated by the addition of ATP through the flash photolysis of P3-1-(2-nitrophenyl)-ethyl adenosine 5'-triphosphate utilizing...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00292a003
更新日期:1983-11-08 00:00:00
abstract::In an effort to extend automated Edman degradation to nanomole quantities of protein, the method of sequenator analysis described by Edman and Begg (Edman, P., and Begg, G. (1967), Eur. J. Biochem. 1, 80) was modified to permit long degradations in the absence of carrier proteins. By using an aqueous 0.1 M Quadrol pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00684a036
更新日期:1975-07-01 00:00:00
abstract::The fungal metabolite gliotoxin has a redox-active disulfide bridge spanning carbons 3 and 6 of a diketopiperazine (DKP) scaffold. The proposed DKP synthetase, GliP, from Aspergillus fumigatus Af293, is a three module (A1-T1-C1-A2-T2-C2-T3) 236 kDa protein that can be overproduced in soluble form in Escherichia coli. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061845b
更新日期:2006-12-19 00:00:00
abstract::The motion of the ethanolamine head group in unsonicated lipid bilayers above and below the phase transition is studied by means of deuterium and phosphorus magnetic resonance. For this purpose, dipalmitoyl-3-sn-phosphatidylethanolamine is selectively deuterated at the two ethanolamine carbon atoms. The deuterium quad...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00669a001
更新日期:1976-11-30 00:00:00
abstract::P-Selectin, expressed on activated endothelial cells and platelets, is a high kinetic adhesion receptor involved in leukocyte rolling of the inflammatory response, or in tumor cell binding in the course of metastasis. Thus, P-selectin inhibition is a promising therapeutic target. The anti-inflammatory and anti-metasta...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi602347g
更新日期:2007-05-22 00:00:00
abstract::NMR spectroscopy is particularly adept at site-specifically monitoring dynamic processes in proteins, such as protein folding, domain movements, ligand binding, and side-chain rotations. By coupling the favorable spectroscopic properties of highly dynamic side-chain methyl groups with transverse-relaxation-optimized s...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/acs.biochem.8b00953
更新日期:2019-01-22 00:00:00
abstract::The binding of the inhibitor 8-nitroxanthine to urate oxidase has been investigated by Raman and UV-visible absorption spectroscopy. The absorption maximum of 8-nitroxanthine shifts from 380 to 400 nm upon binding to the enzyme, demonstrating that the electronic structure of the ligand is perturbed. It has been propos...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0507837
更新日期:2005-08-30 00:00:00
abstract::Calcineurin, a Ca2+, calmodulin-dependent protein phosphatase, was recently found to bind with high affinity to two different immunosuppressant binding proteins (immunophilins) with absolute dependence on the presence of the immunosuppressants FK506 or cyclosporin A (CsA) [Liu et al. (1991) Cell 66, 807-815]. The bind...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00131a002
更新日期:1992-04-28 00:00:00
abstract::F-STOP is a microtubule-associated protein that stabilizes microtubules in a calmodulin (CaM)-dependent manner. All members of the stable tubule only polypeptide (STOP) family have a central domain that contains nearly identical multiple repeats, and a CaM binding motif is present in multiple copies within this domain...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034746w
更新日期:2003-10-07 00:00:00
abstract::The complete amino acid sequences of two basic proline-rich proteins, IB-1 and IB-6, from human parotid saliva have been determined. Fragments for sequence analysis were obtained by enzymatic digestions. The proteins have molecular weights of 9571 (IB-1) and 11,530 (IB-6) and contain 34 and 39 mol % proline, respectiv...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00357a013
更新日期:1986-05-06 00:00:00
abstract::Biotin synthase (BioB) is an iron-sulfur dimeric enzyme which catalyzes the last step in biotin synthesis. The reaction consists of the introduction of a sulfur atom into dethiobiotin. It is shown here that BioB displays a significant cysteine desulfurase activity, providing it with the ability to mobilize sulfur from...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0122011
更新日期:2002-07-23 00:00:00
abstract::Multiple forms of the acute-phase serum protein SAA were isolated from the lipoprotein fraction of plasma from a single individual. These protein forms were purified by size-exclusion, ion-exchange, and reverse-phase high-pressure liquid chromatography, and then the tryptic peptides were subjected to amino acid sequen...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00405a044
更新日期:1988-03-08 00:00:00
abstract::The complex kinetic behavior commonly observed in protein folding studies suggests that a heterogeneous population of molecules exists in solution and that a number of discrete steps are involved in the conversion of unfolded molecules to the fully native form. A central issue in protein folding is whether any of thes...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9700476
更新日期:1997-05-13 00:00:00
abstract::Halorhodopsin from Natronomonas pharaonis (NpHR) acts an inward-directed, light-driven chloride pump and forms a homotrimer. To evaluate effect of trimeric assembly, that is, intermolecular interaction, on the control or modulation of light-driven chloride pumping activity of individual HRs, it is important to underst...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901380c
更新日期:2009-12-29 00:00:00
abstract::Liver alcohol dehydrogenase (LADH) carboxymethylated at Cys-46 (CMLADH) forms two different ternary complexes with 4-trans-(N,N-dimethylamino)cinnamaldehyde (DACA). The complex with reduced nicotinamide adenine dinucleotide (NADH) is characterized by a 38-nm red shift of the long-wavelength pi, pi* transition to 436 n...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00321a085
更新日期:1984-12-18 00:00:00
abstract::The catalysis of the chemical exchange between acetaldehyde and its hydrate by bovine carbonic anhydrase B was investigated by the NMR line broadening and saturation transfer techniques. While both chemical exchange and binding to the enzyme had an effect on the line broadening of acetaldehyde and its hydrate, the sat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00550a021
更新日期:1980-04-29 00:00:00
abstract::Histone deacetylase 8 (HDAC8) is a well-characterized member of the class I acetyl-lysine deacetylase (HDAC) family. Previous work has shown that the efficiency of HDAC8-catalyzed deacetylation of a methylcoumarin peptide varies depending on the identity of the divalent metal ion in the HDAC8 active site. Here we demo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.7b00851
更新日期:2017-10-24 00:00:00
abstract::The interactions of HIV-1 Env (gp120-gp41) with CD4 and coreceptors trigger a barrage of conformational changes in Env that drive the membrane fusion process. Various regions of gp41 have profound effects on HIV entry and budding. However, the precise interactions between gp41 and the membrane have not been elucidated...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701920f
更新日期:2008-02-19 00:00:00
abstract::Post-translational phosphorylation and the related conformational changes in signaling proteins are responsible for regulating a wide range of subcellular processes. Human Pin1 is central to many of these cell signaling pathways in normal and aberrant subcellular processes, catalyzing cis-trans isomerization of the pe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2009954
更新日期:2011-11-08 00:00:00
abstract::N-(1-Pyrenyl)iodoacetamide has been used to introduce fluorescent probes into Escherichia coli RNA polymerase. After an incubation time of 15 min, approximately 2 pyrene equiv was introduced per enzyme molecule. There was no further increase in modification after more extended periods of incubation. Neither calf thymu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00215a028
更新日期:1991-01-08 00:00:00
abstract::Mistic is an unusual membrane protein from Bacillus subtilis. It appears to fold and insert autonomously into a lipid bilayer and has been suggested as a tool that aids the targeting of eukaryotic membrane proteins to bacterial membranes. The NMR structure of Mistic in detergent (LDAO) micelles has revealed it to be a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0608818
更新日期:2006-08-01 00:00:00
abstract::The formation of closed icosahedral capsids from a single species of coat protein subunit requires that the subunits assume different conformations at different lattice positions. In the double-stranded DNA bacteriophage P22, formation of correctly dimensioned capsids is mediated by interaction between coat protein su...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00475a030
更新日期:1990-06-12 00:00:00
abstract::Oxalate is the strongest known inhibitor of yeast cytochrome b2 activity. We have used spectrophotometric titration, temperature-jump relaxation, and calorimetry in an investigation of the interaction between enzyme and inhibitor. The titration data are consistent with noncooperative binding to one site per subunit. T...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00647a002
更新日期:1976-01-27 00:00:00
abstract::The effect of normo (NTG)- and hypertriglyceridemic (HTG)-VLDL on cultured human umbilical vein endothelial cell (HUVEC) surface-localized fibrinolysis was examined following pre-incubation with NTG-, HTG-VLDL, LDL (1-20 micrograms/mL) or buffer (control). Ligand binding assays, using 125I-labeled tcu-PA, t-PA, or Glu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952032i
更新日期:1996-05-14 00:00:00
abstract::Amyloid deposition of human islet amyloid polypeptide (hIAPP) in the islets of Langerhans is closely associated with the pathogenesis of type II diabetes mellitus. Despite substantial evidence linking amyloidogenic hIAPP to loss of β-cell mass and decreased pancreatic function, the molecular mechanism of hIAPP cytotox...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300542g
更新日期:2012-09-04 00:00:00
abstract::Peripheral benzodiazepine receptors (PBR), first described more than 20 years ago, have been attributed with many putative functions including ones in cellular proliferation and cellular respiration. Hence, it is quite conceivable that deregulation of this receptor could lead to pathology. We and others have reported ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi030251v
更新日期:2004-09-28 00:00:00