146-91-8Relevant articles and documents
Mechanistic Insights into the Metal-Dependent Activation of ZnII-Dependent Metallochaperones
Jordan, Matthew R.,Wang, Jiefei,Weiss, Andy,Skaar, Eric P.,Capdevila, Daiana A.,Giedroc, David P.
, p. 13661 - 13672 (2019)
Members of the COG0523 subfamily of candidate GTPase metallochaperones function in bacterial transition-metal homeostasis, but the nature of the cognate metal, mechanism of metal transfer, and identification of target protein(s) for metal delivery remain open questions. Here, we explore the multifunctionality of members of the subfamily linked to delivering ZnII to apoprotein targets under conditions of host-imposed transition-metal depletion. We examine two zinc-uptake repressor (Zur)-regulated COG0523 family members, each from a major human pathogen, Acinetobacter baumannii (AbZigA) and Staphylococcus aureus (SaZigA), in an effort to develop a model for ZnII metallochaperone activity. ZnII chelator competition experiments reveal one high-affinity (KZn1 ≈ 1010-1011 M-1) metal-binding site in each GTPase, while AbZigA and SaZigA are characterized by an additional one and two (lower-affinity) metal-binding sites, respectively. CoII titrations reveal that both metallochaperones have similar electronic absorption characteristics that indicate the presence of two tetrahedral metal coordination sites. High-affinity metal binding at the CXCC motif activates the GTPase activity of both enzymes, with ZnII more effective than CoII. Both GTPases bind the product, GDP, more tightly in the apoprotein than the ZnII-bound state and exhibit what is best described as a "locked" conformation around the GTP substrate. Negative thermodynamic linkage is observed between nucleotide binding and metal binding, leading to a new mechanistic model for COG0523-catalyzed metal delivery.
One-Pot Synthesis of α,γ-Dinucleoside 5'-Triphosphates, G5'pppG and A5'pppA, Using S,S'-Bis(4-chlorophenyl)phosphorodithioate
Fukuoka, Koichiro,Suda, Fuminori,Suzuki, Ryo,Ishikawa, Masahide,Hata, Tsujiaki
, p. 499 - 502 (1994)
S,S'-Bis(4-chlorophenyl) phosphorodithioate was useful for the synthesis of α,γ-dinucleoside 5'-triphosphates, G5'pppG and A5'pppA starting from the corresponding unprotected nucleoside 5'-phosphates under neutral conditions. G5'pppG was used for the synthesis of m7G5'pppG by means of the N7-methylation of one of two guanine moieties of G5'pppG.
Donor substrate binding and enzymatic mechanism of human core α1,6-fucosyltransferase (FUT8)
K?tzler, Miriam P.,Blank, Simon,Bantleon, Frank I.,Spillner, Edzard,Meyer, Bernd
, p. 1915 - 1925 (2012)
Background: Fucosylation is essential for various biological processes including tumorigenesis, inflammation, cell-cell recognition and host-pathogen interactions. Biosynthesis of fucosylated glycans is accomplished by fucosyltransferases. The enzymatic product of core α1,6-fucosyltransferase (FUT8) plays a major role in a plethora of pathological conditions, e.g. in prognosis of hepatocellular carcinoma and in colon cancer. Detailed knowledge of the binding mode of its substrates is required for the design of molecules that can modulate the activity of the enzyme. Methods: We provide a detailed description of binding interactions of human FUT8 with its natural donor substrate GDP-fucose and related compounds. GDP-Fuc was placed in FUT8 by structural analogy to the structure of protein-O-fucosyltransferase (cePOFUT) co-crystallized with GDP-Fuc. The epitope of the donor substrate bound to FUT8 was determined by STD NMR. The in silico model is further supported by experimental data from SPR binding assays. The complex was optimized by molecular dynamics simulations. Results: Guanine is specifically recognized by His363 and Asp453. Furthermore, the pyrophosphate is tightly bound via numerous hydrogen bonds and contributes affinity to a major part. Arg365 was found to bind both the β-phosphate and the fucose moiety at the same time. Conclusions: Discovery of a novel structural analogy between cePOFUT and FUT8 allows the placement of the donor substrate GDP-Fuc. The positioning was confirmed by various experimental and computational techniques. General significance: The model illustrates details of the molecular basis of substrate recognition for a human fucosyltransferase for the first time and, thus, provides a basis for structure-based design of inhibitors.
THE PREPARATION OF ADENOSINE 5'-PYROPHOSPHATE BY A NON-ENZYMIC METHOD.
DAWSON,FORD,EICHBERG
, p. 104 - 106 (1965)
1. A non-enzymic method for the preparation of adenosine 5'-diphosphate
Phosphorylation of ganciclovir phosphonate by cellular GMP kinase determines the stereoselectivity of anti-human cytomegalovirus activity
Miller, Wayne H.,Beauchamp, Lilia M.,Meade, Eric,Reardon, John E.,Biron, Karen K.,Smith, Albert A.,Goss, Charles A.,Miller, Richard L.
, p. 341 - 356 (2000)
A racemic mixture of ganciclovir phosphonate was resolved by stereoselective phosphorylation using GMP kinase. The R-enantiomer of ganciclovir phosphonate was active against human cytomegalovirus but the S- enantiomer was less active. We show that enantiomeric selectivity of antiviral activity for ganciclovir phosphonate was conferred by stereoselective phosphorylations by mammalian enzymes, not by stereoslective inhibition of DNA polymerase from human cytomegalovirus.
Functionally nonequivalent interactions of guanosine 5'-triphosphate, inosine 5'-triphosphate, and xanthosine 5'-triphosphate with the retinal G-protein, transducin, and with G(i)-proteins in HL-60 leukemia cell membranes
Klinker, Jan F.,Seifert, Roland
, p. 551 - 562 (1997)
G proteins mediate signal transfer from receptors to effector systems. In their guanosine 5'-triphosphate (GTP) bound form, G-protein α-subunits activate effector systems. Termination of G-protein activation is achieved by the high-affinity GTPase [E.C. 3.6.1.-] of their α-subunits. Like GTP, inosine 5' -triphosphate (ITP) and xanthosine 5' triphosphate (XTP) can support effector system activation. We studied the interactions of GTP, ITP, and XTP with the retinal G protein, transducin (TD), and with G-proteins in HL-60 leukemia cell membranes. TD hydrolyzed nucleoside 5'-triphosphates (NTPs) in the order of efficacy GTP > ITP > XTP. NTPs eluted TD from rod outer segment disk membranes in the same order of efficacy. ITP and XTP competitively inhibited TD catalyzed GTP hydrolysis. In HL-60 membranes, the chemoattractants N-formyl-L-methionyl-L-leucyl-L- phenylaline (fMLP) and leukotriene B4 (LTB4,) effectively activated GTP and ITP hydrolysis by G(i) proteins. fMLP and LTB4, were at least l0-fold more potent activators of ITPase than of GTPasc. Complement C5a effectively activated the GTPase of G(i)-proteins but was only a weak stimulator of ITPase. The potency of C5a to activate GTP and ITP hydrolysis was similar. The fMLP stimulated GTPase had a lower K(m) value than the fMLP-stimulated ITPase, whereas the opposite was true for the V(max) values. fMLP, C5a, and LTB4 did not stimulate XTP hydrolysis. Collectively, our data show that GTP, ITP, and XTP bind to G-proteins with different affinities, that G-proteins hydrolyze NTPs with different efficacies, and that chemoattractants stimulate GTP and ITP hydrolysis by G(i)-proteins in a receptor-specific manner. On the basis of our results and the data in the literature, we put forward the hypothesis that GTP, ITP, and XTP act as differential signal amplifiers and signal sorters at the G-protein level.
Biochemical properties of the human guanylate binding protein 5 and a tumor-specific truncated splice variant
Wehner, Mark,Herrmann, Christian
, p. 1597 - 1605 (2010)
The human guanylate binding protein 5 (hGBP5) belongs to the family of interferon-γ-inducible large GTPases, which are well known for their high induction by pro-inflammatory cytokines. The cellular role of this protein family is unclear at this point, but there are indications for antiviral and antibacterial activity of hGBP1. hGBP5 exists in three splice variants, forming two different proteins, of which the tumor-specific one is C-terminally truncated by 97 amino acids, and therefore lacks the CaaX motif for geranylgeranylation. Here we present biochemical data on the splice variants of hGBP5. We show that, unlike hGBP1, hGBP5a/b and hGBP5ta do not bind GMP or produce any GMP during hydrolysis despite the fact the residues involved in GMP production from hGBP1 are conserved in hGBP5. Hydrolysis of GTP is concentration-dependent and shows weak self-activation. Thermodynamic studies showed strongly negative entropic changes during nucleotide binding, which reflect structural ordering in the protein during nucleotide binding. These structural changes were also observed during changes in the oligomerization state. We observed only a minor influence of the C-terminal truncation on hydrolysis, nucleotide binding and oligomerization of hGBP5. Based on these similarities we speculate that the missing C-terminal part, which also carries the geranylgeranylation motif, is the reason for the dysregulation of hGBP5′s function in lymphoma cells.
Enzymatic synthesis of UTPγS, a potent hydrolysis resistant agonist of P2U-purinoceptors
Lazarowski, Eduardo R.,Watt, William C.,Stutts, M. Jackson,Brown, H. Alex,Boucher, Richard C.,Harden, T. Kendall
, p. 203 - 209 (1996)
1 The defective Cl- secretion characteristic of cystic fibrosis airway epithelial cells can be bypassed by an alternative Ca2+ dependent Cl- secretory pathway that is activated by extracellular nucleotides, e.g. uridine-5′triphosphate (UTP), acting on P2U purinoceptors. Since UTP is susceptible to hydrolysis by nucleotidases and phosphatases present in the airways, the identification of stable P2U-purinoceptor agonists would be of therapeutic relevance. 2 Uridine-5′-O-(3-thiotriphosphate) (UTPγS) was synthesized by nucleoside diphosphate kinase-catalyzed transfer of the γ-phosphorothioate from guanosine-5′-O-(3-thiotriphosphate) (GTPγS) or adenosine-5′-O-(3-thiotriphosphate) (ATPγS) to UDP. Formation of UTPγS was illustrated by observation of transfer of 35S from [35S]-GTPγS and transfer of 3H from [3H]-UDP. The chemical identity of high performance liquid chromatography (h.p.l.c.)-purified UTPγS was confirmed by nuclear magnetic resonance analysis. 3 Human 1321N1 astrocytoma cells stably expressing the phospholipase C-coupled human P2U-purinoceptor were utilized to lest the activity of UTPγS. UTPγS (EC50 = 240 nM) was essentially equipotent to UTP and ATP for stimulation of inositol phosphate formation. 4 Unlike [3H]-UTP, [3H]-UTPγS was not hydrolyzed by alkaline phosphatase, acid phosphatase, or apyrase. Moreover, no hydrolysis was detected during a 1 h incubation with human nasal epithelial cells. 5 UTPγS was equally potent and efficacious with UTP for stimulation of Cl- secretion by human nasal epithelium from both normal donors and cystic fibrosis patients. Based on its high potency and resistance to hydrolysis, UTPγS represents a promising compound for treatment of cystic fibrosis.
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Gibson et al.
, p. 86,88 (1956)
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Helicobacter hepaticus Hh0072 gene encodes a novel α1-3- fucosyltransferase belonging to CAZy GT11 family
Zhang, Lei,Lau, Kam,Cheng, Jiansong,Yu, Hai,Li, Yanhong,Sugiarto, Go,Huang, Shengshu,Ding, Li,Thon, Vireak,Wang, Peng G,Chen, Xi
, p. 1077 - 1088 (2010)
Lewis x (Lex) and sialyl Lewis x (SLex)-containing glycans play important roles in numerous physiological and pathological processes. The key enzyme for the final step formation of these Lewis antigens is α1-3-fucosyltransferase. Here we report molecular cloning and functional expression of a novel Helicobacter hepaticus α1-3- fucosyltransferase (HhFT1) which shows activity towards both non-sialylated and sialylated Type II oligosaccharide acceptor substrates. It is a promising catalyst for enzymatic and chemoenzymatic synthesis of Lex, sialyl Lex and their derivatives. Unlike all other α1-3/4- fucosyltransferases characterized so far which belong to Carbohydrate Active Enzyme (CAZy, http://www.cazy.org/) glycosyltransferase family GT10, the HhFT1 shares protein sequence homology with α1-2-fucosyltransferases and belongs to CAZy glycosyltransferase family GT11. The HhFT1 is thus the first α1-3-fucosyltransferase identified in the GT11 family.
Photo-electrochemical Bioanalysis of Guanosine Monophosphate Using Coupled Enzymatic Reactions at a CdS/ZnS Quantum Dot Electrode
Sabir, Nadeem,Khan, Nazimuddin,V?lkner, Johannes,Widdascheck, Felix,Del Pino, Pablo,Witte, Gregor,Riedel, Marc,Lisdat, Fred,Konrad, Manfred,Parak, Wolfgang J.
, p. 5844 - 5850 (2015)
A photo-electrochemical sensor for the specific detection of guanosine monophosphate (GMP) is demonstrated, based on three enzymes combined in a coupled reaction assay. The first reaction involves the adenosine triphosphate (ATP)-dependent conversion of GMP to guanosine diphosphate (GDP) by guanylate kinase, which warrants substrate specificity. The reaction products ADP and GDPare co-substrates for the enzymatic conversion of phosphoenolpyruvate to pyruvate in a second reaction mediated by pyruvate kinase. Pyruvate in turn is the co-substrate for lactate dehydrogenase that generates lactate via oxidation of nicotinamide adenine dinucleotide (reduced form) NADH to NAD+. This third enzymatic reaction is electrochemically detected. For this purpose a CdS/ZnS quantum dot (QD) electrode is illuminated and the photocurrent response under fixed potential conditions is evaluated. The sequential enzyme reactions are first evaluated in solution. Subsequently, a sensor for GMP is constructed using polyelectrolytes for enzyme immobilization.
Borate-nucleotide complex formation depends on charge and phosphorylation state
Kim, Danny H.,Faull, Kym F.,Norris, Andrew J.,Eckhert, Curtis D.
, p. 743 - 751 (2004)
Flow injection analysis with electrospray ionization mass spectrometry was used to investigate borate-nucleotide complex formation. Solutions containing 100 μM nucleotide and 500 μM boric acid in water-acetonitrile-triethylamine (50:50:0.2, v/v/v; pH 10.3
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Kotelnikova,Dovedova
, p. 594 (1954)
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