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Multiple N-Acetyltransferases and Drug Metabolism TISSUE DISTRIBUTION, CHARACTERIZATION and SIGNIFICANCE of MAMMALIAN N-ACETYLTRANSFERASE by D
Biochem. J. (1973) 132, 519-526 519 Printed in Great Britain Multiple N-Acetyltransferases and Drug Metabolism TISSUE DISTRIBUTION, CHARACTERIZATION AND SIGNIFICANCE OF MAMMALIAN N-ACETYLTRANSFERASE By D. J. HEARSE* and W. W. WEBER Department ofPharmacology, New York University Medical Center, 550 First Avenue, New York, N. Y. 10016, U.S.A. (Received 6 October 1972) Investigations in the rabbit have indicated the existence of more than one N-acetyl- transferase (EC 2.3.1.5). At least two enzymes, possibly isoenzymes, were partially characterized. The enzymes differed in their tissue distribution, substrate specificity, stability and pH characteristics. One of the enzymes was primarily associated with liver and gut and catalysed the acetylation of a wide range of drugs and foreign compounds, e.g. isoniazid, p-aminobenzoic acid, sulphamethazine and sulphadiazine. The activity of this enzyme corresponded to the well-characterized polymorphic trait of isoniazid acetylation, and determined whether individuals were classified as either 'rapid' or 'slow' acetylators. Another enzyme activity found in extrahepatic tissues readily catalysed the acetylation ofp-aminobenzoic acid but was much less active towards isoniazid and sulpha- methazine. The activity of this enzyme remained relatively constant from individual to individual. Studies in vitro and in vivo with both 'rapid' and 'slow' acetylator rabbits re- vealed that, for certain substrates, extrahepatic N-acetyltransferase contributes signifi- cantly to the total acetylating capacity of the individual. The possible significance and applicability ofthese findings to drugmetabolism and acetylation polymorphism in man is discussed. Liver N-acetyltransferase catalyses the acetylation purified by the same procedure and their pH charac- of a number of commonly used drugs and foreign teristics, heat stabilities, kinetic properties, substrate compounds such as isoniazid, sulphamethazine, specificities and reaction mechanisms are indis- sulphadiazine, p-aminobenzoic acid, diamino- tinguishable. -
1 Metabolic Dysfunction Is Restricted to the Sciatic Nerve in Experimental
Page 1 of 255 Diabetes Metabolic dysfunction is restricted to the sciatic nerve in experimental diabetic neuropathy Oliver J. Freeman1,2, Richard D. Unwin2,3, Andrew W. Dowsey2,3, Paul Begley2,3, Sumia Ali1, Katherine A. Hollywood2,3, Nitin Rustogi2,3, Rasmus S. Petersen1, Warwick B. Dunn2,3†, Garth J.S. Cooper2,3,4,5* & Natalie J. Gardiner1* 1 Faculty of Life Sciences, University of Manchester, UK 2 Centre for Advanced Discovery and Experimental Therapeutics (CADET), Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Sciences Centre, Manchester, UK 3 Centre for Endocrinology and Diabetes, Institute of Human Development, Faculty of Medical and Human Sciences, University of Manchester, UK 4 School of Biological Sciences, University of Auckland, New Zealand 5 Department of Pharmacology, Medical Sciences Division, University of Oxford, UK † Present address: School of Biosciences, University of Birmingham, UK *Joint corresponding authors: Natalie J. Gardiner and Garth J.S. Cooper Email: [email protected]; [email protected] Address: University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, United Kingdom Telephone: +44 161 275 5768; +44 161 701 0240 Word count: 4,490 Number of tables: 1, Number of figures: 6 Running title: Metabolic dysfunction in diabetic neuropathy 1 Diabetes Publish Ahead of Print, published online October 15, 2015 Diabetes Page 2 of 255 Abstract High glucose levels in the peripheral nervous system (PNS) have been implicated in the pathogenesis of diabetic neuropathy (DN). However our understanding of the molecular mechanisms which cause the marked distal pathology is incomplete. Here we performed a comprehensive, system-wide analysis of the PNS of a rodent model of DN. -
KAT5 Acetylates Cgas to Promote Innate Immune Response to DNA Virus
KAT5 acetylates cGAS to promote innate immune response to DNA virus Ze-Min Songa, Heng Lina, Xue-Mei Yia, Wei Guoa, Ming-Ming Hua, and Hong-Bing Shua,1 aDepartment of Infectious Diseases, Zhongnan Hospital of Wuhan University, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Wuhan University, 430071 Wuhan, China Edited by Adolfo Garcia-Sastre, Icahn School of Medicine at Mount Sinai, New York, NY, and approved July 30, 2020 (received for review December 19, 2019) The DNA sensor cGMP-AMP synthase (cGAS) senses cytosolic mi- suppress its enzymatic activity (15). It has also been shown that crobial or self DNA to initiate a MITA/STING-dependent innate im- the NUD of cGAS is critically involved in its optimal DNA- mune response. cGAS is regulated by various posttranslational binding (16), phase-separation (7), and subcellular locations modifications at its C-terminal catalytic domain. Whether and (17). However, whether and how the NUD of cGAS is regulated how its N-terminal unstructured domain is regulated by posttrans- remains unknown. lational modifications remain unknown. We identified the acetyl- The lysine acetyltransferase 5 (KAT5) is a catalytic subunit of transferase KAT5 as a positive regulator of cGAS-mediated innate the highly conserved NuA4 acetyltransferase complex, which immune signaling. Overexpression of KAT5 potentiated viral- plays critical roles in DNA damage repair, p53-mediated apo- DNA–triggered transcription of downstream antiviral genes, whereas ptosis, HIV-1 transcription, and autophagy (18–21). Although a KAT5 deficiency had the opposite effects. Mice with inactivated KAT5 has been investigated mostly as a transcriptional regula- Kat5 exhibited lower levels of serum cytokines in response to DNA tor, there is increasing evidence that KAT5 also acts as a key virus infection, higher viral titers in the brains, and more susceptibility regulator in signal transduction pathways by targeting nonhis- to DNA-virus–induced death. -
Ancestral Class-Promiscuity As a Driver of Functional Diversity in the BAHD
bioRxiv preprint doi: https://doi.org/10.1101/2020.11.18.385815; this version posted November 20, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Ancestral class-promiscuity as a driver of functional diversity in the 2 BAHD acyltransferase family in plants 3 Lars H. Kruse1, Austin T. Weigle3, Jesús Martínez-Gómez1,2, Jason D. Chobirko1,5, Jason 4 E. Schaffer6, Alexandra A. Bennett1,7, Chelsea D. Specht1,2, Joseph M. Jez6, Diwakar 5 Shukla4, Gaurav D. Moghe1* 6 Footnotes: 7 1 Plant Biology Section, School of Integrative Plant Sciences, Cornell University, Ithaca, 8 NY, 14853, USA 9 2 L.H. Bailey Hortorium, Cornell University, Ithaca, NY, 14853, USA 10 3 Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, 11 USA 12 4 Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana- 13 Champaign, Urbana, IL, 61801, USA 14 5 Present address: Department of Molecular Biology and Genetics, Cornell University, 15 Ithaca, NY, 14853, USA 16 6 Department of Biology, Washington University in St. Louis, St. Louis, MO, 63130, USA 17 7 Present address: Institute of Analytical Chemistry, Universität für Bodenkultur Wien, 18 Vienna, 1190, Austria 19 20 * Corresponding author: [email protected] 21 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.11.18.385815; this version posted November 20, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. -
Supp Table 6.Pdf
Supplementary Table 6. Processes associated to the 2037 SCL candidate target genes ID Symbol Entrez Gene Name Process NM_178114 AMIGO2 adhesion molecule with Ig-like domain 2 adhesion NM_033474 ARVCF armadillo repeat gene deletes in velocardiofacial syndrome adhesion NM_027060 BTBD9 BTB (POZ) domain containing 9 adhesion NM_001039149 CD226 CD226 molecule adhesion NM_010581 CD47 CD47 molecule adhesion NM_023370 CDH23 cadherin-like 23 adhesion NM_207298 CERCAM cerebral endothelial cell adhesion molecule adhesion NM_021719 CLDN15 claudin 15 adhesion NM_009902 CLDN3 claudin 3 adhesion NM_008779 CNTN3 contactin 3 (plasmacytoma associated) adhesion NM_015734 COL5A1 collagen, type V, alpha 1 adhesion NM_007803 CTTN cortactin adhesion NM_009142 CX3CL1 chemokine (C-X3-C motif) ligand 1 adhesion NM_031174 DSCAM Down syndrome cell adhesion molecule adhesion NM_145158 EMILIN2 elastin microfibril interfacer 2 adhesion NM_001081286 FAT1 FAT tumor suppressor homolog 1 (Drosophila) adhesion NM_001080814 FAT3 FAT tumor suppressor homolog 3 (Drosophila) adhesion NM_153795 FERMT3 fermitin family homolog 3 (Drosophila) adhesion NM_010494 ICAM2 intercellular adhesion molecule 2 adhesion NM_023892 ICAM4 (includes EG:3386) intercellular adhesion molecule 4 (Landsteiner-Wiener blood group)adhesion NM_001001979 MEGF10 multiple EGF-like-domains 10 adhesion NM_172522 MEGF11 multiple EGF-like-domains 11 adhesion NM_010739 MUC13 mucin 13, cell surface associated adhesion NM_013610 NINJ1 ninjurin 1 adhesion NM_016718 NINJ2 ninjurin 2 adhesion NM_172932 NLGN3 neuroligin -
TIP55, a Splice Isoform of the KAT5 Acetyltransferase, Is
www.nature.com/scientificreports OPEN TIP55, a splice isoform of the KAT5 acetyltransferase, is essential for developmental gene regulation and Received: 27 March 2018 Accepted: 24 September 2018 organogenesis Published: xx xx xxxx Diwash Acharya1, Bernadette Nera1, Zachary J. Milstone2,3, Lauren Bourke 2,3, Yeonsoo Yoon4, Jaime A. Rivera-Pérez4, Chinmay M. Trivedi 1,2,3 & Thomas G. Fazzio1 Regulation of chromatin structure is critical for cell type-specifc gene expression. Many chromatin regulatory complexes exist in several diferent forms, due to alternative splicing and diferential incorporation of accessory subunits. However, in vivo studies often utilize mutations that eliminate multiple forms of complexes, preventing assessment of the specifc roles of each. Here we examined the developmental roles of the TIP55 isoform of the KAT5 histone acetyltransferase. In contrast to the pre-implantation lethal phenotype of mice lacking all four Kat5 transcripts, mice specifcally defcient for Tip55 die around embryonic day 11.5 (E11.5). Prior to developmental arrest, defects in heart and neural tube were evident in Tip55 mutant embryos. Specifcation of cardiac and neural cell fates appeared normal in Tip55 mutants. However, cell division and survival were impaired in heart and neural tube, respectively, revealing a role for TIP55 in cellular proliferation. Consistent with these fndings, transcriptome profling revealed perturbations in genes that function in multiple cell types and developmental pathways. These fndings show that Tip55 is dispensable for the pre- and early post-implantation roles of Kat5, but is essential during organogenesis. Our results raise the possibility that isoform-specifc functions of other chromatin regulatory proteins may play important roles in development. -
Haem Oxygenase Is Synthetically Lethal with the Tumour Suppressor Fumarate Hydratase
LETTER doi:10.1038/nature10363 Haem oxygenase is synthetically lethal with the tumour suppressor fumarate hydratase Christian Frezza1, Liang Zheng1, Ori Folger2, Kartik N. Rajagopalan3, Elaine D. MacKenzie1, Livnat Jerby2, Massimo Micaroni4, Barbara Chaneton1, Julie Adam5, Ann Hedley1, Gabriela Kalna1, Ian P. M. Tomlinson6, Patrick J. Pollard5, Dave G. Watson7, Ralph J. Deberardinis3, Tomer Shlomi8*, Eytan Ruppin2,9* & Eyal Gottlieb1 Fumarate hydratase (FH) is an enzyme of the tricarboxylic acid majority of fumarate was unlabelled (m10), indicating that glucose is cycle (TCA cycle) that catalyses the hydration of fumarate into a minor source of carbon for the TCA cycle in both Fh1fl/fl and Fh12/2 malate. Germline mutations of FH are responsible for hereditary cells (Fig. 1e). On the other hand, when cells were incubated with 13C- leiomyomatosis and renal-cell cancer (HLRCC)1. It has previously glutamine most of the fumarate was labelled (Fig. 1f). In Fh12/2 cells been demonstrated that the absence of FH leads to the accumula- cultured with uniformly labelled glutamine (Fig. 1f), the vast majority of tion of fumarate, which activates hypoxia-inducible factors (HIFs) the labelled fumarate contained all four carbon atoms derived from at normal oxygen tensions2–4. However, so far no mechanism that glutamine (m14). By contrast, in Fh1fl/fl cells, the fumarate pool con- explains the ability of cells to survive without a functional TCA tained substantial fractions of molecules with fewer than four 13C atoms cycle has been provided. Here we use newly characterized genetically due to processing of fumarate beyond the Fh1 step. The lack of these modified kidney mouse cells in which Fh1 has been deleted, and products in Fh12/2 cells confirms that no accessory pathway exists in apply a newly developed computer model of the metabolism of these these cells to circumvent the loss of Fh1 enzymatic activity, indicating a cells to predict and experimentally validate a linear metabolic path- true blockade of the cycle. -
©Ferrata Storti Foundation
Original Articles T-cell/histiocyte-rich large B-cell lymphoma shows transcriptional features suggestive of a tolerogenic host immune response Peter Van Loo,1,2,3 Thomas Tousseyn,4 Vera Vanhentenrijk,4 Daan Dierickx,5 Agnieszka Malecka,6 Isabelle Vanden Bempt,4 Gregor Verhoef,5 Jan Delabie,6 Peter Marynen,1,2 Patrick Matthys,7 and Chris De Wolf-Peeters4 1Department of Molecular and Developmental Genetics, VIB, Leuven, Belgium; 2Department of Human Genetics, K.U.Leuven, Leuven, Belgium; 3Bioinformatics Group, Department of Electrical Engineering, K.U.Leuven, Leuven, Belgium; 4Department of Pathology, University Hospitals K.U.Leuven, Leuven, Belgium; 5Department of Hematology, University Hospitals K.U.Leuven, Leuven, Belgium; 6Department of Pathology, The Norwegian Radium Hospital, University of Oslo, Oslo, Norway, and 7Department of Microbiology and Immunology, Rega Institute for Medical Research, K.U.Leuven, Leuven, Belgium Citation: Van Loo P, Tousseyn T, Vanhentenrijk V, Dierickx D, Malecka A, Vanden Bempt I, Verhoef G, Delabie J, Marynen P, Matthys P, and De Wolf-Peeters C. T-cell/histiocyte-rich large B-cell lymphoma shows transcriptional features suggestive of a tolero- genic host immune response. Haematologica. 2010;95:440-448. doi:10.3324/haematol.2009.009647 The Online Supplementary Tables S1-5 are in separate PDF files Supplementary Design and Methods One microgram of total RNA was reverse transcribed using random primers and SuperScript II (Invitrogen, Merelbeke, Validation of microarray results by real-time quantitative Belgium), as recommended by the manufacturer. Relative reverse transcriptase polymerase chain reaction quantification was subsequently performed using the compar- Ten genes measured by microarray gene expression profil- ative CT method (see User Bulletin #2: Relative Quantitation ing were validated by real-time quantitative reverse transcrip- of Gene Expression, Applied Biosystems). -
Recombinant Human Biliverdin Reductase B/BLVRB
Recombinant Human Biliverdin Reductase B/BLVRB Catalog Number: 6568-BR DESCRIPTION Source E. coliderived human Biliverdin Reductase B/BLVRB protein Ala2Gln206, with an Nterminal Met and 6His tag Accession # P30043 Nterminal Sequence Nterminus confirmed by detection of His tag using Western analysis. Analysis Predicted Molecular 23 kDa Mass SPECIFICATIONS SDSPAGE 2325 kDa, reducing conditions Activity Measured by the reduction of riboflavin 5'monophosphate (FMN) using NADPH as the cofactor. The specific activity is >225 pmol/min/μg, as measured under the described conditions. Endotoxin Level <1.0 EU per 1 μg of the protein by the LAL method. Purity >95%, by SDSPAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Brij and Glycerol. See Certificate of Analysis for details. Activity Assay Protocol Materials l Assay Buffer: 100 mM Sodium Acetate, pH 5.0 l Recombinant Human Biliverdin Reductase B/BLVRB (rhBLVRB) (Catalog # 6568BR) l Riboflavin 5’monophosphate sodium salt dihydrate (FMN) (Sigma, Catalog # F6750), 10 mM in deionized water l βNicotinamide adenine dinucleotide phosphate reduced, tetrasodium salt (βNADPH) (Sigma, Catalog # N7505), 10 mM in deionized water l UV Plate (Costar, Catalog # 3635) l Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent Assay 1. Dilute rhBLVRB to 40 ng/μL in Assay Buffer. 2. Prepare a Reaction Mixture by combining FMN and βNADPH in Assay Buffer to a concentration of 400 μM for each. 3. Load 50 μL of 40 ng/μL rhBLVRB into the microplate, and start the reaction by adding 50 μL of Reaction Mixture. -
Chemistry of Acetyl Transfer by Histone Modifying Enzymes: Structure, Mechanism and Implications for Effector Design
Oncogene (2007) 26, 5528–5540 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc REVIEW Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design SC Hodawadekar and R Marmorstein The Wistar Institute and The Department of Chemistry, University of Pennsylvania, Philadelphia, PA, USA The post-translational modification of histones plays an remodeling proteins that mobilize the histone proteins important role in chromatin regulation, a process that within chromatin (Varga-Weisz and Becker, 2006); insures the fidelity of gene expression and other DNA histone chaperone proteins that assemble, disassemble transactions. Of the enzymes that mediate post-transla- or replace variant histones within chromatin (Loyola tion modification, the histone acetyltransferase (HAT) and Almouzni, 2004); and post-translational modifica- and histone deacetylase (HDAC) proteins that add and tion enzymes that add or remove functional groups to or remove acetyl groups to and from target lysine residues from the histone proteins (Nightingale et al., 2006). within histones, respectively, have been the most exten- The post-translational modifications of histones sively studied at both the functional and structural levels. involve the addition or removal of acetyl, methyl or Not surprisingly, the aberrant activity of several of these phosphate groups as well as the reversible transfer of the enzymes have been implicated in human diseases such as ubiquitin and sumo proteins. -
Kidney V-Atpase-Rich Cell Proteome Database
A comprehensive list of the proteins that are expressed in V-ATPase-rich cells harvested from the kidneys based on the isolation by enzymatic digestion and fluorescence-activated cell sorting (FACS) from transgenic B1-EGFP mice, which express EGFP under the control of the promoter of the V-ATPase-B1 subunit. In these mice, type A and B intercalated cells and connecting segment principal cells of the kidney express EGFP. The protein identification was performed by LC-MS/MS using an LTQ tandem mass spectrometer (Thermo Fisher Scientific). For questions or comments please contact Sylvie Breton ([email protected]) or Mark A. Knepper ([email protected]). -
Impact of the Genes UGT1A1, GSTT1, GSTM1, GSTA1, GSTP1 and NAT2 on Acute Alcohol-Toxic Hepatitis
Cent. Eur. J. Biol. • 9(2) • 2014 • 125-130 DOI: 10.2478/s11535-013-0249-y Central European Journal of Biology Impact of the genes UGT1A1, GSTT1, GSTM1, GSTA1, GSTP1 and NAT2 on acute alcohol-toxic hepatitis Research Article Linda Piekuse1*, Baiba Lace2, Madara Kreile1, Lilite Sadovska2, Inga Kempa1, Zanda Daneberga3, Ieva Mičule3, Valentina Sondore4, Jazeps Keiss4, Astrida Krumina2 1Scientific Laboratory of Molecular Genetics Riga Stradins University, 1007 Riga, Latvia 2Latvian Biomedical Research and Study Center, 1067 Riga, Latvia 3Medical Genetics Clinic, University Children’s Hospital, 1004 Riga, Latvia 4Latvian Centre of Infectious Diseases, Riga East University Hospital, 1006 Riga, Latvia Received 27 March 2013; Accepted 04 August 2013 Abstract: Alcohol metabolism causes cellular damage by changing the redox status of cells. In this study, we investigated the relationship between genetic markers in genes coding for enzymes involved in cellular redox stabilization and their potential role in the clinical outcome of acute alcohol-induced hepatitis. Study subjects comprised 60 patients with acute alcohol-induced hepatitis. The control group consisted of 122 healthy non-related individuals. Eight genetic markers of the genes UGT1A1, GSTA1, GSTP1, NAT2, GSTT1 and GSTM1 were genotyped. GSTT1 null genotype was identified as a risk allele for alcohol-toxic hepatitis progression (OR 2.146, P=0.013). It was also found to correlate negatively with the level of prothrombin (β= –11.05, P=0.037) and positively with hyaluronic acid (β=170.4, P=0.014). NAT2 gene alleles rs1799929 and rs1799930 showed opposing associations with the activity of the biochemical markers γ-glutamyltransferase and alkaline phosphatase; rs1799929 was negatively correlated with γ-glutamyltransferase (β=–261.3, P=0.018) and alkaline phosphatase (β= –270.5, P=0.032), whereas rs1799930 was positively correlated with γ-glutamyltransferase (β=325.8, P=0.011) and alkaline phosphatase (β=374.8, P=0.011).