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1.
Although several investigations have focused on luminescence modulation by chelation with metal cations using bidentate ligands or crown ether systems, a bis(crown ether) system has not yet been used for modulation of chemiluminescence (CL) reactions. In the CL reaction of 2-(phenyl and 4-dimethylaminophenyl)-4-hydroperoxy-4-3',4'-(15-crown-5)phenyl-5-3',4'(15-crown-5)phenyl-4H-isoimidazoles 2a and b possessing a bis(15-crown-5 ether) moiety, the rate acceleration was observed in the presence of K(+), Rb(+) and Cs(+) due to the holding effect of the bis-crown moiety, but no rate acceleration was observed by Li(+) and Na(+) due to the template effect of the crown moiety. The acceleration of the CL reaction rates is ascribable to the conformational change induced by the scissor-like motion of the bis-crown moiety assisted by the holding effect. 相似文献
2.
T Ueda Y Isakari H Aoki T Yasukochi S Masutomo K Kawano Y Terada H Yamada T Imoto 《Journal of biochemistry》1991,109(5):690-698
We prepared the lysozyme derivative in which the beta-carboxyl group of Asp101 was modified with alpha-O-methyl N-glycylglucosaminide as an amide by means of the carbodimide reaction (alpha-MGG lysozyme). Since Asp101 residue is located at the edge of the active site cleft, a 1H-NMR study was carried out for this derivative in order to investigate the interaction between the introduced substituent and the active site cleft. It was confirmed that the alpha-MGG moiety sat in the active site cleft in alpha-MGG lysozyme from the reduction of line broadening of the NH-proton of Trp63 located in the active site cleft, the remarkable chemical shift change of the methyl group of the alpha-MGG moiety upon adding a trimer of N-acetyl-D-glucosamine [(NAG)3], and the NOE between the C6-proton resonance of Trp63 and the methyl resonance of the alpha-MGG moiety. Furthermore, alpha-MGG lysozyme had increased thermal stability compared with native lysozyme. Therefore, it was concluded that the alpha-MGG moiety covalently attached to Asp101 interacted with the active site cleft to increase the thermal stability of lysozyme. 相似文献
3.
Synthetic Sialyl Compounds as Well as Natural Gangliosides Induce Neuritogenesis in a Mouse Neuroblastoma Cell Line (Neuro2a) 总被引:13,自引:9,他引:4
Shuichi Tsuji Tatsuro Yamashita Makoto Tanaka Yoshitaka Nagai† 《Journal of neurochemistry》1988,50(2):414-423
Amphipathic compounds containing N-acetylneuraminic acid (sialic acid) [for example, D-N-acetylneuraminyl-(alpha 2-1)-2S,3R,4E-2-N-tetracosanoyl sphingenine, sialyl alkyl glycerol ethers, and sialyl cholesterols] induced neuritogenesis in a neuroblastoma cell line (Neuro2a). The sialic acid in the hydrophilic moiety of the compounds is specifically required for neuritogenesis. The requirement for molecular specificity of the hydrophobic moiety, however, is rather low. Regarding the hydrophobic moiety, no preference for cholesterol, alkyl glycerol ether, or ceramide residues was observed as to their neuritogenic activity. Sialyl compounds with alpha-ketosidic sialyl linkages were more active than the corresponding beta-anomers. These sialyl compounds induced the growth of only one neurite, but a long one, from the cell body. This type of neuritogenes is completely different from that induced by compounds capable of elevating the concentration of intracellular cyclic AMP, which induced the appearance of many neurites from a single cell body. Besides this morphological change, the active sialyl compounds also caused a change in the carbohydrate composition of the cell surface. Sialyl compound treatment drastically increased the concentration of peanut agglutinin binding sites. 相似文献
4.
The thermodynamics of cofactor binding to the isolated reductase domain (Red) of nNOS and its mutants have been studied by isothermal titration calorimetry. The NADP(+) and 2',5'-ADP binding stoichiometry to Red were both 1:1, consistent with a one-site kinetic model instead of a two-site model. The binding constant (K(D) = 71 nM) and the large heat capacity change (ΔC(p) = -440 cal mol(-1) K(-1)) for 2',5'-ADP were remarkably different from those for NADP(+) (1.7 μM and -140 cal mol(-1) K(-1), respectively). These results indicate that the nicotinamide moiety as well as the adenosine moiety has an important role in binding to nNOS. They also suggest that the thermodynamics of the conformational change in Red caused by cofactor binding are significantly different from the conformational changes that occur in cytochrome c reductase, in which the nicotinamide moiety of the cofactor is not essential for binding. Analysis of the deletion mutant of the autoinhibitory helix (RedΔ40) revealed that the deletion resulted in a decrease in the binding affinity of 2',5'-ADP with more unfavorable enthalpy gain. In the case of RedCaM, which contains a calmodulin (CaM) binding site, the presence of Ca(2+)/CaM caused a 6.7-fold increase in the binding affinity for 2',5'-ADP that was mostly due to the favorable entropy change. These results are consistent with a model in which Ca(2+)/CaM induces a conformational change in NOS to a flexible "open" form from a "closed" form that locked by cofactor binding, and this change facilitates the electron transfer required for catalysis. 相似文献
5.
Iwaki S Hanaoka K Piao W Komatsu T Ueno T Terai T Nagano T 《Bioorganic & medicinal chemistry letters》2012,22(8):2798-2802
Hypoxia occurs in various diseases, including cancer, ischemia, and acute and chronic vascular diseases. Here we describe the design and synthesis of the first hypoxia-sensitive MRI contrast agents, SAGds. SAGds showed a pH-dependent r(1) relaxivity change associated with intramolecular chelation of the nitrogen atom of the sulfonamide moiety to the Gd(3+) center. There was a correlation between the pK(a) of the r(1) relaxivity change and the sum of the Hammett σ constants of substituents on the aromatic ring. Among the synthesized compounds, 4NO(2)2MeOSAGd was selectively reduced to the amine by rat liver microsomes under hypoxic conditions, resulting in a 1.8-fold increment of the r(1) relaxivity owing to the change in pK(a) of the arylsulfonamide moiety. This enhancement of the r(1) relaxivity could be clearly detected in T(1)-weighted MR images. Thus, 4NO(2)2MeOSAGd is a 'smart' MRI contrast agent for the detection of hypoxia under physiological conditions. 相似文献
6.
A convenient synthesis of labelled rhodopsin and studies on its active site 总被引:3,自引:2,他引:1
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Digitonin solutions of labelled rhodopsin, containing (3)H in the retinyl moiety, were prepared by two related methods. Labelled rhodopsin was also prepared for the first time in cetyltrimethylammonium bromide and purified by column chromatography. It was shown that only certain rhodopsin preparations on denaturation in the dark and the reduction with sodium borohydride gave up to 60% of the radioactivity in a fraction characterized as N-retinylphosphatidylethanolamine. Such preparations also gave a lipid-linked retinyl moiety at the metarhodopsin-I stage, but, as expected, a protein-linked retinyl moiety at the metarhodopsin-II stage. Other preparations however, gave exclusively protein-bound radioactivity at the native-rhodopsin, metarhodopsin-I and metarhodopsin-II stages. It is therefore conceivable that the formation of N-retinylphosphatidylethanolamine is due to a non-enzymic reaction resulting from the transfer of the retinyl moiety from its native site to an amino group of a favourably oriented phospholipid molecule. The only firmly established aspect of the rhodopsin active site remains the demonstration in our previous work that at the metarhodopsin-II stage the retinyl moiety is linked to an in-amino group of lysine. On the basis of chemical reactivity it is argued that the light-induced conversion of rhodopsin into metarhodopsin II involves a profound conformational change resulting in the dislocation of the retinylideneiminium chromophore from a non-polar environment in rhodopsin to a polar environment in metarhodopsin II. 相似文献
7.
利用H+-ATP酶复合体(也称ATP合成酶)中的Fo的色氨酸荧光,观察了复合体中F1结合ATP或ADP(酶蛋白与底物分子比为1:1)时,Fo的荧光猝灭常数的变化(用竹红菌乙作为膜区蛋白荧光的猝灭剂)结果表明F1结合ATP或ADP时Fo可得到不同的猝灭常数(Ksv),也就是Fo会产生不同的构象变化。加入二价金属离子起动ATP水解反应结束后:ATP+H2O→ADP+Pi,这时可以在Fo观察到与ADP加Mg2+时相同猝灭常数Ksv;用荧光强度随时间进程变化的实验可观察到F1水解过程中导致Fo构象变化的动力学过程。这些结果说明了H+-ATP酶复合体ATP合成的过程中F1与Fo之间存在着构象之间的通信与传递。 相似文献
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10.
《Nucleosides, nucleotides & nucleic acids》2013,32(10-11):1751-1760
We have prepared four diastereomers of TT dinucleotides containing an isoxazoline moiety and four dodecanucleotides incorporating these TT dinucleotides in the middle of the DNA sequence. We also have determined the melting temperatures of these modified oligonucleotides (Tm values) by measuring change in UV absorbance. 相似文献
11.
During the catalytic cycle of beta1,4-galactosyltransferase-1 (Gal-T1), upon the binding of Mn(2+) followed by UDP-Gal, two flexible loops, a long and a short loop, change their conformation from open to closed. We have determined the crystal structures of a human M340H-Gal-T1 mutant in the open conformation (apo-enzyme), its Mn(2+) and Mn(2+)-UDP-Gal-bound complexes, and of a pentenary complex of bovine Gal-T1-Mn(2+)-UDP-GalNAc-Glc-alpha-lactalbumin. These studies show that during the conformational changes in Gal-T1, the coordination of Mn(2+) undergoes significant changes. It loses a coordination bond with a water molecule bound in the open conformation of Gal-T1 while forming a new coordination bond with another water molecule in the closed conformation, creating an active ground-state structure that facilitates enzyme catalysis. In the crystal structure of the pentenary complex, the N-acetylglucosamine (GlcNAc) moiety is found cleaved from UDP-GalNAc and is placed 2.7A away from the O4 oxygen atom of the acceptor Glc molecule, yet to form the product. The anomeric C1 atom of the cleaved GalNAc moiety has only two covalent bonds with its non-hydrogen atoms (O5 and C2 atoms), similar to either an oxocarbenium ion or N-acetylgalactal form, which are crystallographically indistinguishable at the present resolution. The structure also shows that the newly formed, metal-coordinating water molecule forms a hydrogen bond with the beta-phosphate group of the cleaved UDP moiety. This hydrogen bond formation results in the rotation of the beta-phosphate group of UDP away from the cleaved GalNAc moiety, thereby preventing the re-formation of the UDP-sugar during catalysis. Therefore, this water molecule plays an important role during catalysis in ensuring that the catalytic reaction proceeds in a forward direction. 相似文献
12.
A series of 17 hydrochlorides of piperidinylpropyl esters of alkoxy-substituted phenylcarbamic acids with the alkoxy group in position 2, 3 or 4 on the phenyl ring, and basic substituents attached to the moiety in position 3, were evaluated for in vitro antimycobacterial activity against the strains of Mycobacterium tuberculosis, M. kansasii and M. avium. To describe the structure-antimycobacterial activity relationships (QSAR), an approach based on the Free-Wilson method was employed to express the differences between individual moieties (including propyl and ethyl). The change of ethyl to propyl moiety increases the activity to M. tuberculosis but decreases the antimycobacterial activity to all potentially pathogenic strains under study. 相似文献
13.
《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2017,1862(11):1414-1423
Bacterial lipoproteins are a subset of membrane proteins localized on either leaflet of the lipid bilayer. These proteins are anchored to membranes through their N-terminal lipid moiety attached to a conserved Cys. Since the protein moiety of most lipoproteins is hydrophilic, they are expected to play various roles in a hydrophilic environment outside the cytoplasmic membrane. Gram-negative bacteria such as Escherichia coli possess an outer membrane, to which most lipoproteins are sorted. The Lol pathway plays a central role in the sorting of lipoproteins to the outer membrane after lipoprotein precursors are processed to mature forms in the cytoplasmic membrane. Most lipoproteins are anchored to the inner leaflet of the outer membrane with their protein moiety in the periplasm. However, recent studies indicated that some lipoproteins further undergo topology change in the outer membrane, and play critical roles in the biogenesis and quality control of the outer membrane.This article is part of a Special Issue entitled: Bacterial Lipids edited by Russell E. Bishop. 相似文献
14.
The use of antibody molecules in immunoassay, molecular targeting, or detection techniques encompasses a broad variety of applications affecting nearly every field of medical science. In cancer therapy, monoclonal antibodies (mAb) have been used as vehicles to deliver radionuclides, toxins, or drugs to the target cancer cells. New conjugation methods are most needed to conjugate a wide variety of targeting small molecules and peptidomimatic compounds. Here, we exploited a keto-oxime method for conjugation of protease susceptible linkers to an antibody. This modified method involves two steps: (i) introduction of methyl ketone linkers (referred to as linker moiety) to the primary amines present in the antibody and (ii) conjugation of ketone linkers to aminoxy functional group present in the conjugated moiety (referred to as functional moiety). We have optimized this conjugation method and shown that approximately 10 functional moieties can be conjugated to antibody. Conjugation was verified by MALDI-TOF MS and Western blot analysis. The acidic pH conditions used in this method did not change the immune reactivity of the Ab. In addition, in vitro protease susceptibility assay was performed to validate this method for prodrug release assay as well as to remove excess radioimmune conjugates from circulation. This orthogonal method is compatible with peptides containing a thiol, amino, or carboxyl groups in the conjugation moiety. 相似文献
15.
The single tryptophan residue, at position 69 in the amino acid sequence, was used as an intrinsic probe to obtain structural and dynamical information on the lipolytic enzyme Fusarium solani cutinase. In the enzyme's native state the tryptophan fluorescence is highly quenched. Time-resolved experiments reveal that the majority of the excited state species is characterized by an unusually fast decay time of approximately 40 ps, indicating the occurrence of a very efficient nonradiative relaxation process, possibly via the adjacent disulphide bond or via the peptide bonds of a nearby loop. A minority of the excited state species relaxes on a nanosecond time scale. Irradiation of the enzyme in the tryptophan absorption band causes an increase by an order of magnitude of the fluorescence quantum yield. This increase is ascribed to a photo-induced, subtle structural change of a minor subset of species whose fluorescence is not highly quenched. The structural change is accompanied by a tightening of the local environment of the tryptophan moiety, as indicated by results from time-resolved fluorescence anisotropy which reveal a complete disappearance of the segmental flexibility of the tryptophan moiety. 相似文献
16.
Exo-1,3-beta-D-glucanase secreted by Saccharomyces cerevisiae undergoes extracellular modifications in its carbohydrate moiety that change the affinity towards the lectin concanavalin A. The transition of negatively reacting enzyme form into positively reacting one depends on temperature. Results from experiments with glucono-delta-lactone and from treatments in vitro with hydrolases suggest a glycosidase-mediated mechanism. 相似文献
17.
The appearance of positive charge on the nitrogen moiety of phthalideisoquinoline alkaloids brings about a strong change in the intensities of CD Cotton effects. Hence the optical rotations of enantiomeric salts and their parent base of identical configuration are often of opposite sign. Thus, the name (+)-bicuculline-methiodide for the methiodide salt of (+)-bicuculline is false. 相似文献
18.
Caffeic acid phenethyl ester (CAPE) is a polyphenolic natural product that possesses numerous biological activities including anti-inflammatory effects. CAPE-mediated nuclear factor-erythroid 2 p45 (NF-E2)-related factor 2 (Nrf2) activation is likely responsible for some of its biological effects. CAPE was chemically modified to yield CAPE analogues that were subjected to experiments examining cellular Nrf2 activity. CAPE and the CAPE analogue with a catechol moiety, but not the other analogues, activated the Nrf2 pathway. In addition, only biotin-labeled CAPE analogues with the catechol moiety precipitated Kelch-like ECH associated protein 1 (Keap1) when incubated with cell lysates and streptavidin agarose beads. Sodium hypochlorite (NaOCl) oxidation of the catechol moiety in CAPE produced an oxidized, electrophilic form of CAPE (Oxi-CAPE) and greatly enhanced the ability of CAPE to activate Nrf2 and to bind to Keap1. Rectal administration of CAPE ameliorated 2,4,6-trinitrobenzene sulfonic acid-induced rat colitis and activated the Nrf2 pathway in the inflamed colon, and incubation of CAPE in the lumen of the inflamed distal colon generated Oxi-CAPE. However, these biological effects and chemical change of CAPE were not observed in the normal colon. Our data suggest that CAPE requires the catechol moiety for the oxidation-enhanced activation of the Nrf2 pathway and has potential as a pathologically targeted Nrf2-activating agent that is exclusively activated in pathological states with oxidative stress such as colonic inflammation. 相似文献
19.
Osamu Kurahashi Kenzo Yokozeki Shigeru Yamanaka Hitoshi Enei 《Bioscience, biotechnology, and biochemistry》2013,77(12):3211-3212
Antihypertensive compounds were purified from an extract of autologous Lactobacillus casei cell lysates. The most effective compounds were polysaccharide-glycopeptide complexes, found in the cell wall. The average molecular weight was estimated as 180,000 from gel filtration using Sephacryl S-300. The polysaccharide moiety of the complexes consisted of glucose, rhamnose, and galactose, whereas the glycopeptide moiety consisted of N-acetylglucosamine, N-acetylmuramic acid, asparagine, glutamine, alanine, and lysine. The varieties of the components of these moieties were constant and independent of complex molecular size. When these complexes were orally administered to spontaneously hypertensive rats (SHR) and renal hypertensive rats (RHR) at doses of 1 mg/kg-body weight, systolic blood pressure (SBP) decreased by 10–20 mmHg 6 to 12 hr after administration without any change in heart rate. Appreciable hypotensive activity was lost by treating the complexes with hydrofluoric acid, which hydrolytically cleaves the phosphodiester bond between the polysaccharide and glycopeptide moiety. 相似文献
20.
Alpha-aminoadipate aminotransferase (AAA-AT), a homolog of mammalian kynurenine aminotransferase II (Kat II), transfers an amino group to 2-oxoadipate to yield alpha-aminoadipate in lysine biosynthesis through the alpha-aminoadipate pathway in Thermus thermophilus. AAA-AT catalyzes transamination against various substrates, including AAA, glutamate, leucine, and aromatic amino acids. To elucidate the structural change for recognition of various substrates, we determined crystal structures of AAA-AT in four forms: with pyridoxal 5'-phosphate (PLP) (PLP complex), with PLP and leucine (PLP/Leu complex), with N-phosphopyridoxyl-leucine (PPL) (PPL complex), and with N-phosphopyridoxyl-alpha-aminoadipate (PPA) at 2.67, 2.26, 1.75, and 1.67 A resolution, respectively. The PLP complex is in an open state, whereas PLP/Leu, PPL, and PPA complexes are in closed states with maximal displacement (over 7 A) of the alpha2 helix and the beta1 strand in the small domain to cover the active site, indicating that conformational change is induced by substrate binding. In PPL and PLP/Leu complexes, several hydrophobic residues on the alpha2 helix recognize the hydrophobic side chain of the bound leucine moiety whereas, in the PPA complex, the alpha2 helix rotates to place the guanidium moiety of Arg23 on the helix at the appropriate position to interact with the carboxyl side chain of the AAA moiety. These results indicate that AAA-AT can recognize various kinds of substrates using the mobile alpha2 helix. The crystal structures and site-directed mutagenesis revealed that intersubunit-electrostatic interactions contribute to the elevated thermostability of this enzyme. 相似文献