Abstract:
:Accumulation of the cytotoxic 40- to 42-residue beta-amyloid peptide represents the primary pathological process in Alzheimer's disease (AD). BACE1 (beta-site APP cleaving enzyme 1) is responsible for the initial required step in the neuronal amyloidogenic processing of beta-amyloid precursor protein and is a major drug target for the therapeutic intervention of AD. In the present study, BACE1 is initially synthesized as an immature precursor protein containing part of the pre domain and the entire pro domain, and undergoes autocatalytic conversion to yield the well-folded mature BACE1 enzyme. To understand the mechanism of the conversion and the role of the pro domain, we monitored the autocatalytic conversion of BACE1 by heteronuclear NMR spectroscopy and used chemical shift perturbations as a probe to study the structural changes accompanying the autocatalytic conversion. NMR data revealed local conformational changes from a partially disordered to a well-folded conformation associated with the conversion. The conformational changes are largely concentrated in the NH(2)-terminal lobe. Conversely, the active site conformations are conserved during the autocatalytic conversion. The precursor and mature BACE1 proteins were further characterized for their ability to interact with a substrate-based transition state BACE1 peptide inhibitor. The precursor BACE1 rapidly adopted the bound conformation in the presence of the inhibitor, which is identical to the bound conformation of the mature protein. The interaction of the inhibitor with both the precursor BACE1 and the fully processed BACE1 is in slow exchange on the NMR time scale, indicating a tight binding interaction. Overall, the NMR data demonstrated that the pro domain does not hinder inhibitor binding and may assist in the proper folding of the protein. The fully processed BACE1 represents a high quality well-folded protein which is highly stable over a long period of time, and is suitable for evaluation of inhibitor binding by NMR for drug intervention.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Wang YS,Beyer BM,Senior MM,Wyss DFdoi
10.1021/bi051040osubject
Has Abstractpub_date
2005-12-20 00:00:00pages
16594-601issue
50eissn
0006-2960issn
1520-4995journal_volume
44pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Kinetics of reduction of phototrophic bacterial flavocytochromes c by exogenous flavin semiquinones and fully reduced flavins generated by laser flash photolysis have been studied. The mechanisms of reduction of Chromatium and Chlorobium flavocytochromes c are more similar to one another than previously thought. Neith...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00327a002
更新日期:1985-03-12 00:00:00
abstract::Human lung tryptase (HLT), a trypsin-like serine proteinase stored as an active enzyme in association with heparin in mast cell granules, is released into the extracellular environment when mast cells are activated. Tryptases are unusual in that they form tetramers and bind heparin. As there are no known endogenous tr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960042t
更新日期:1996-10-22 00:00:00
abstract::In an attempt to understand the mechanism by which transcription factors compete with histone octamers for cognate binding sites in chromatin, the effect of the histone binding protein nucleoplasmin on the binding of TFIIIA to nucleosomal 5S rRNA genes was tested. In this study, it was shown that, despite the previous...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972723+
更新日期:1998-02-03 00:00:00
abstract::The stereoselectivity of the adenosine cyclic 3',5'-phosphate (cAMP) binding sites on the regulatory subunit of the type II bovine cardiac muscle cAMP-dependent protein kinase was investigated by examining the interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates (cAMPS) with these sites. While activ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00261a028
更新日期:1982-08-31 00:00:00
abstract::FKBP12, a small human enzyme, aids protein folding by catalyzing cis-trans isomerization of peptidyl-prolyl bonds, and is involved in cell signaling pathways, calcium regulation, and the immune response. The underlying molecular mechanisms are not fully understood, but it is well-known that aromatic residues in the ac...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b01157
更新日期:2017-01-10 00:00:00
abstract::S-Glutathionylation is a redox-regulated modification that uncouples endothelial nitric oxide synthase (eNOS), switching its function from nitric oxide (NO) synthesis to (•)O2(-) generation, and serves to regulate vascular function. While in vitro or in vivo eNOS S-glutathionylation with modification of Cys689 and Cys...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400404s
更新日期:2013-09-24 00:00:00
abstract::The phosphorylation mechanism of Dictyostelium discoideum nucleoside diphosphate (NDP) kinase was investigated by NMR. 31P chemical shifts were measured on both native and denatured enzyme. In the enzymatically phosphorylated enzyme denatured by 9 M urea or 7 M guanidine hydrochloride, the NDP kinase phosphohistidine ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00038a043
更新日期:1995-09-26 00:00:00
abstract::Alpha2u-globulin is a male rat liver protein under multihormonal control which represents approximately 1% of hepatic protein synthesis. We have measured the rate of transcription of the alpha2u-globulin gene using a nuclear cell-free transcriptional system with mercurated CTP as substrate for the endogenous RNA polym...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00608a023
更新日期:1978-07-25 00:00:00
abstract::Trimethylamine dehydrogenase (TMADH) from the bacterium Methylophilus methylotrophus (sp. W(3)A(1)) and its C30A mutant were inactivated with three known inactivators of monoamine oxidase, namely, phenylhydrazine, N-cyclopropyl-alpha-methylbenzylamine, and 1-phenylcyclopropylamine. All three compounds irreversibly ina...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0105126
更新日期:2001-07-24 00:00:00
abstract::Thermal transitions of type I collagen fibrils were investigated by differential scanning calorimetry and spectrophotometry of turbidity within a wide range of external conditions. The advanced microcalorimeter allowed us to carry out the measurements at low concentrations of collagen (0.15-0.3 mg/mL). At these concen...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980360n
更新日期:1998-06-02 00:00:00
abstract::Histidine decarboxylase (HDC) from Lactobacillus 30a forms a trimer around a central cavity or well. Three active sites are formed around the well at the interface of each of two adjacent molecules. HDC exhibits cooperative kinetics at pH 7.6 and can be described in terms of a two-state, T and R, model. At pH 4.8, pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00018a009
更新日期:1995-05-09 00:00:00
abstract::When the single-stranded RNA genome of HIV-1 is copied into double-stranded DNA, the viral enzyme reverse transcriptase (RT) catalyzes the addition of approximately 20,000 nucleotides; however, the precise mechanism of nucleotide addition is unknown. In this study, we attempt to integrate the genetic data and biochemi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00016a006
更新日期:1995-04-25 00:00:00
abstract::Membrane-bound phosphodiesterase 6 (PDE6) plays an important role in visual signal transduction by regulating cGMP levels in rod photoreceptor cells. Our understanding of PDE6 catalysis and structure suffers from inadequate characterization of the α and β subunit catalytic core, interactions of the core with two intri...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2014695
更新日期:2011-11-08 00:00:00
abstract::It is known that the overall charge of a protein can change as the molecule approaches a charged object like another protein or a cell membrane. We have formalized this mechanism using a statistical mechanical framework and show how this rather overlooked interaction increases the attraction between protein molecules....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047630o
更新日期:2005-04-19 00:00:00
abstract::Structure formation and dimerization are concerted processes in the refolding of Arc repressor. The integrity of secondary structure in the transition state of Arc refolding has been investigated here by determining the changes in equilibrium stability and refolding/unfolding kinetics for a set of Ala --> Gly mutation...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000423d
更新日期:2000-07-18 00:00:00
abstract::Multimodular enzymes, including polyketide synthases (PKSs), nonribosomal peptide synthetases (NRPSs), and mixed PKS/NRPS systems, contain functional domains with similar functions. Domain swapping and module fusion are potential powerful strategies for creating hybrid enzymes to synthesize modified natural products. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047538s
更新日期:2005-03-29 00:00:00
abstract::Fluorescein isothiocyanate derivatization of the human lactotransferrin on Lys-264 inhibits the binding of the protein of human PHA-activated lymphocytes [Legrand, D., Mazurier, J., Maes, P., Rochard, E., Montreuil, J., & Spik, G. (1991) Biochem. J. 276, 733-738], indicating that part of the receptor-binding site is l...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00153a018
更新日期:1992-09-29 00:00:00
abstract::The interactions of two plant annexins, annexin 24(Ca32) from Capsicum annuum and annexin Gh1 from Gossypium hirsutum, with phospholipid membranes have been characterized using liposome-based assays and adsorption to monolayers. These two plant annexins show a preference for phosphatidylserine-containing membranes and...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0516226
更新日期:2005-12-13 00:00:00
abstract::Nitrophorins 1-4 (NP1-4) are ferriheme proteins from the blood-sucking insect Rhodnius prolixus that transport nitric oxide (NO) to the victim, sequester histamine, and inhibit blood coagulation. Here, we report kinetic and thermodynamic analyses for ligand binding by all four proteins and their reduction potentials. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000766b
更新日期:2000-08-22 00:00:00
abstract::Static FTIR light-induced difference spectra have been recorded for reaction centers from Rhodopseudomonas viridis in the following charge-separated states: P+QA(-)-PQA, P+QB(-)-PQB, I(-)-I, I-QA(-)-IQA, and I-QA(2-)-IQA. A comparison of the I(-)-I difference spectra with the I-QA(-)-IQA difference spectra reveals new...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00120a006
更新日期:1992-02-11 00:00:00
abstract::Alpha-synuclein modulates dopamine homeostasis in dopamine-producing neurons of substantia nigra, partly through regulation of human dopamine transporter (hDAT) activity. To identify the underlying mechanisms, we disrupted the modulation of hDAT activity by wild-type (wt) alpha-synuclein, and its familial Parkinson's ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035308s
更新日期:2004-02-10 00:00:00
abstract::A fluorescence method is presented for quantitatively analyzing exocytosis phenomena and monitoring their kinetics. The method is based on the particular properties of a hydrophobic fluorescent probe, 1-[4-(trimethylammonio)phenyl]-6-phenylhexa-1,3,5-triene (TMA-DPH) [Prendergast, F.G., Haugland, R.P., & Callahan, P.J...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00356a045
更新日期:1986-04-22 00:00:00
abstract::Biogenesis of pyrroloquinoline quinone (PQQ) in Klebsiella pneumoniae requires the expression of six genes (pqqA-F). One of these genes (pqqE) encodes a 43 kDa protein (PqqE) that plays a role in the initial steps in PQQ formation [Veletrop, J. S., et al. (1995) J. Bacteriol. 177, 5088-5098]. PqqE contains two highly ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi900918b
更新日期:2009-10-27 00:00:00
abstract::Mitomycins make up a group of antitumor natural products that are biosynthesized from aminohydroxybenzoic acid (AHBA) and N-acetylglucosamine (GlcNAc). While the biosynthetic gene cluster was reported two decades ago, the mechanism by which the two building blocks, AHBA and GlcNAc, are coupled during biosynthesis rema...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.9b00379
更新日期:2019-06-25 00:00:00
abstract::Gemcitabine 5'-diphosphate (F(2)CDP) is a potent inhibitor of ribonucleotide reductases (RNRs), enzymes that convert nucleotides (NDPs) to deoxynucleotides and are essential for DNA replication and repair. The Escherichia coli RNR, an alpha2beta2 complex, when incubated with 1 equiv of F(2)CDP catalyzes the release of...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901590q
更新日期:2009-12-15 00:00:00
abstract::Cholera toxin, an activator of adenylate cyclase in a wide variety of cells, is a substrate for the phosphotransferase reaction catalyzed by purified cyclic adenosine 5'-monophosphate dependent bovine cardiac muscle protein kinase and the protein associated with human erythrocyte membranes. Phosphorylation occurs when...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00658a031
更新日期:1976-06-29 00:00:00
abstract::1,3-15N-Labeled uracil was synthesized chemically and used to prepare labeled Escherichia coli tRNA(Val) biosynthetically. 500-MHz measurements of 15N and proton chemical shift were obtained, for all uridine and uridine-related bases, by heteronuclear multiple-quantum coherence spectroscopy. All the uracil NH group re...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00166a013
更新日期:1992-12-29 00:00:00
abstract::We have purified the junctional acetylcholine receptor from normal rat skeletal muscle and compared its structure with that of the extrajunctional receptor from denervated muscle. The two receptors from leg muscle were distinguished by isoelectric focusing and by reaction with sera from patients with myasthenia gravis...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00582a028
更新日期:1979-07-24 00:00:00
abstract::We found that recombinant human adult hemoglobin (rHb A) expressed in Escherichia coli showed heterogeneity of components with the intensity of a positive CD band at 260 nm and that it could be resolved into three components (SP-1, SP-2, and SP-3) by SP-Sepharose column chromatography. 1H NMR revealed that SP-1 is ide...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7015519
更新日期:2008-01-15 00:00:00
abstract::The phosphocarrier protein HPr of the phosphoenolpyruvate-dependent sugar transport system of Escherichia coli can exist in a phosphorylated and a nonphosphorylated form. During phosphorylation, the phosphoryl group is carried on a histidine residue, His15. The hydrogen-bonding state of this histidine was examined wit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00487a026
更新日期:1990-09-04 00:00:00