Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:6.2.1.1 (ACS)
 78,556 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We present a method providing synchronized measurements using the two techniques: quartz crystal microbalance with dissipation (QCM-D) monitoring and localized surface plasmon resonance (LSPR). This was achieved by letting a thin gold film perforated with short-ranged ordered plasmon-active nanoholes act as one of the electrodes of a QCM-D crystal. This enabled transmission-mode optical spectroscopy to be used to temporally resolve colorimetric changes of the LSPR active substrate induced upon biomolecular binding events. The LSPR response could thus be compared with simultaneously obtained changes in resonance frequency, Deltaf, and energy dissipation, DeltaD, of the QCM-D device. Since the LSPR technique is preferentially sensitive to changes within the voids of the nanoholes, while the QCM-D technique is preferentially sensitive to reactions on the planar region between the holes, a surface chemistry providing the same binding kinetics on both gold and silica was used. This was achieved by coating the substrate with poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), which was shown to bind in the same manner on silica and gold modified with a carboxyl-terminated thiol. In this way, the combined setup provided new information about structural changes upon PLL-g-PEG adsorption. We also demonstrate subsequent binding of NeutrAvidin and an immunoreaction utilizing biotin-modified IgG. The combined information from the synchronized measurements was also used in a new way to estimate the sensing volume of the LSPR sensor.
ACS Nano 2008 Oct 28
PMID:Synchronized quartz crystal microbalance and nanoplasmonic sensing of biomolecular recognition reactions. 1920 65

Aqueous dispersible detonation nanodiamonds (NDs) with a diameter of 2-8 nm were assembled into a closely packed ND multilayer nanofilm with positively charged poly-L-lysine via the layer-by-layer deposition technique. The innate biocompatibility of the NDs in both free-floating and thin-film forms was confirmed via cellular gene expression examination by real-time polymerase chain reaction as well as MTT and DNA fragmentation assays. The highly biologically amenable ND nanofilm was successfully integrated with therapeutic molecules, and the functionality of the composite drug-ND material was assessed via interrogation of the suppression of inflammatory cytokine release. Knockdown of lipopolysaccharide-mediated inflammation was observed through the potent attenuation of tumor necrosis factor-alpha, interleukin-6, and inducible nitric oxide synthase levels following ND nanofilm interfacing with RAW 264.7 murine macrophages. Furthermore, basal cytokine secretion levels were assessed to examine innate material biocompability, revealing unchanged cellular inflammatory responses which strongly supported the relevance of the NDs as effective treatment platforms for nanoscale medicine. In addition to the easy preparation, robustness, and fine controllability of the film structures, these hybrid materials possess enormous potential for biomedical applications such as localized drug delivery and anti-inflammatory implant coatings and devices, as demonstrated in vitro in this work.
ACS Nano 2008 Feb
PMID:Protein-mediated assembly of nanodiamond hydrogels into a biocompatible and biofunctional multilayer nanofilm. 1920 20

The use of fixed charge nanopores in practical applications requires tuning externally the electrostatic interaction between the charged groups and the ionic permeants in order to allow integrating a variety of functions on the same nanostructure. We design, produce, and characterize, theoretically and experimentally, a single-track amphoteric nanopore functionalized with lysine and histidine chains whose positive and negative charges are very sensitive to the external pH. This nanofluidic diode with amphoteric chains attached to the pore surface allows for a broad set of rectification properties supported by a single nanodevice. A definite plus is to functionalize these groups on a conical nanopore with well-defined, controlled structural asymmetry which gives virtually every rectification characteristic that may be required in practical applications. Nanometerscaled amphoteric pores are of general interest because of the potential applications in drug delivery systems, ion-exchange membranes for separation of biomacromolecules, antifouling materials with reduced molecular adsorption, and biochemical sensors.
ACS Nano 2009 Mar 24
PMID:A pH-tunable nanofluidic diode with a broad range of rectifying properties. 1922 30

Methods that introduce posttranslational modifications in a general, mild, and non-sequence-specific manner using biologically produced peptides have great utility for investigation of the functions of these modifications. In this study, the substrate promiscuity of a lantibiotic synthetase was exploited for the preparation of phosphopeptides, glycopeptides, and peptides containing analogs of methylated or acetylated lysine residues. Peptides attached to the C-terminus of the leader peptide of the lacticin 481 precursor peptide were phosphorylated on serine residues in a wide variety of sequence contexts by the R399M and T405A mutants of lacticin 481 synthetase (LctM). Serine residues located as many as 30 amino acids C-terminal to the leader peptide were phosphorylated. Wild-type LctM was shown to dehydrate these peptides to generate dehydroalanine-containing products that can be conveniently modified with external nucleophiles including thiosaccharides, 2-(dimethylamino)ethanethiol, and N-acetyl cysteamine, resulting in mimics of O-linked glycopeptides and acetylated and methylated lysines.
ACS Chem Biol 2009 May 15
PMID:Lacticin 481 synthetase as a general serine/threonine kinase. 1929 52

Natural polyphenols with previously demonstrated anticancer potential, epigallocatechin gallate (EGCG), tannic acid, curcumin, and theaflavin, were encased into gelatin-based 200 nm nanoparticles consisting of a soft gel-like interior with or without a surrounding LbL shell of polyelectrolytes (polystyrene sulfonate/polyallylamine hydrochloride, polyglutamic acid/poly-l-lysine, dextran sulfate/protamine sulfate, carboxymethyl cellulose/gelatin, type A) assembled using the layer-by-layer technique. The characteristics of polyphenol loading and factors affecting their release from the nanocapsules were investigated. Nanoparticle-encapsulated EGCG retained its biological activity and blocked hepatocyte growth factor (HGF)-induced intracellular signaling in the breast cancer cell line MBA-MD-231 as potently as free EGCG.
ACS Nano 2009 Jul 28
PMID:Layer-by-Layer-Coated Gelatin Nanoparticles as a Vehicle for Delivery of Natural Polyphenols. 1953 72

SIRT3 is a major mitochondrial NAD(+)-dependent protein deacetylase playing important roles in regulating mitochondrial metabolism and energy production and has been linked to the beneficial effects of exercise and caloric restriction. SIRT3 is emerging as a potential therapeutic target to treat metabolic and neurological diseases. We report the first sets of crystal structures of human SIRT3, an apo-structure with no substrate, a structure with a peptide containing acetyl lysine of its natural substrate acetyl-CoA synthetase 2, a reaction intermediate structure trapped by a thioacetyl peptide, and a structure with the dethioacetylated peptide bound. These structures provide insights into the conformational changes induced by the two substrates required for the reaction, the acetylated substrate peptide and NAD(+). In addition, the binding study by isothermal titration calorimetry suggests that the acetylated peptide is the first substrate to bind to SIRT3, before NAD(+). These structures and biophysical studies provide key insight into the structural and functional relationship of the SIRT3 deacetylation activity.
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PMID:Crystal structures of human SIRT3 displaying substrate-induced conformational changes. 1953 40

Post-translational modification of histones plays an integral role in regulation of genomic expression through modulation of chromatin structure and function. Chemical preparations of histones bearing these modifications allows for comprehensive in vitro mechanistic investigation into their action to deconvolute observations from genome-wide studies in vivo. Previously, we reported the semisynthesis of ubiquitylated histone H2B (uH2B) using two orthogonal expressed protein ligation reactions. Semisynthetic uH2B, when incorporated into nucleosomes, directly stimulates methylation of histone H3 lysine 79 (K79) by the methyltransferase, disruptor of telomeric silencing-like (Dot1L). Although recruitment of Dot1L to the nucleosomal surface by uH2B could be excluded, comprehensive mechanistic analysis was precluded by systematic limitations in the ability to generate uH2B in large scale. Here we report a highly optimized synthesis of ubiquitylated H2B bearing a G76A point mutation u(G76A)H2B, yielding tens of milligrams of ubiquitylated protein. u(G76A)H2B is indistinguishable from the native uH2B by Dot1L, allowing for detailed studies of the resultant trans-histone crosstalk. Kinetic and structure-activity relationship analyses using u(G76A)H2B suggest a noncanonical role for ubiquitin in the enhancement of the chemical step of H3K79 methylation. Furthermore, titration of the level of uH2B within the nucleosome revealed a 1:1 stoichiometry of Dot1L activation.
ACS Chem Biol 2009 Nov 20
PMID:Structure-activity analysis of semisynthetic nucleosomes: mechanistic insights into the stimulation of Dot1L by ubiquitylated histone H2B. 1979 66

Recently, a flow-based selectin-dependent method for the capture and enrichment of specific types of cells (CD34+ hematopoetic stem and progenitor cells and human leukemia HL60) from peripheral blood was demonstrated. However, these devices depend on a monolayer of selectin protein, which has been shown to have a maximum binding efficiency as a function of surface area. A novel surface coating of colloidal silica nanoparticles was designed that alters the surface roughness resulting in increased surface area. The nanoparticles were adhered using either an inorganic titanate resinous coating or an organic polymer of poly-L-lysine. Using Alexa Fluor 647 conjugated P-selectin, an increase in protein adsorption of up to 35% when compared to control was observed. During perfusion experiments using P-selectin-coated microtubes, we observed increased cell capture and greatly decreased rolling velocity at equivalent protein concentration compared to nonparticle control. Atomic force microscopy showed increased surface roughness consistent with the nanoparticle mean diameter, suggesting a monolayer of particles. These results support the coating's potential to improve existing cell capture implantable devices for a variety of therapeutic and scientific uses.
ACS Nano 2010 Jan 26
PMID:Nanoparticle coatings for enhanced capture of flowing cells in microtubes. 2001 20

Sirtuins are NAD(+)-dependent protein deacetylases. They mediate adaptive responses to a variety of stresses, including calorie restriction and metabolic stress. Sirtuin 3 (SIRT3) is localized in the mitochondrial matrix, where it regulates the acetylation levels of metabolic enzymes, including acetyl coenzyme A synthetase 2 (refs 1, 2). Mice lacking both Sirt3 alleles appear phenotypically normal under basal conditions, but show marked hyperacetylation of several mitochondrial proteins. Here we report that SIRT3 expression is upregulated during fasting in liver and brown adipose tissues. During fasting, livers from mice lacking SIRT3 had higher levels of fatty-acid oxidation intermediate products and triglycerides, associated with decreased levels of fatty-acid oxidation, compared to livers from wild-type mice. Mass spectrometry of mitochondrial proteins shows that long-chain acyl coenzyme A dehydrogenase (LCAD) is hyperacetylated at lysine 42 in the absence of SIRT3. LCAD is deacetylated in wild-type mice under fasted conditions and by SIRT3 in vitro and in vivo; and hyperacetylation of LCAD reduces its enzymatic activity. Mice lacking SIRT3 exhibit hallmarks of fatty-acid oxidation disorders during fasting, including reduced ATP levels and intolerance to cold exposure. These findings identify acetylation as a novel regulatory mechanism for mitochondrial fatty-acid oxidation and demonstrate that SIRT3 modulates mitochondrial intermediary metabolism and fatty-acid use during fasting.
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PMID:SIRT3 regulates mitochondrial fatty-acid oxidation by reversible enzyme deacetylation. 2020 11

Spontaneous embedding of gold nanoparticle (NP) aggregates or polyelectrolyte microcapsules modified with NPs in biocompatible hyaluronic acid/poly(l-lysine) films is reported. The NPs were adsorbed in the aggregated state to induce near-IR light absorption. The films functionalized with gold NPs become active in response to a "biologically friendly" near-IR laser at a power of about 20 mW. The activation is characterized by a localized temperature increase in the film, allowing conversion of light energy to heat into confined volumes. Microcapsules adsorbed onto the film can release its cargo under stimulation with near-IR light because of localized permeability changes in their walls. This work is aimed at layer-by-layer film-based biomedical coatings and active surfaces with light-sensitive features wherein metal NPs and microcapsules are used as active centers or carriers with remote control of functionalities.
ACS Appl Mater Interfaces 2009 Aug
PMID:Remote near-IR light activation of a hyaluronic acid/poly(l-lysine) multilayered film and film-entrapped microcapsules. 2035 86


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