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Carbamylation

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https://www.readbyqxmd.com/read/28442530/breach-of-autoreactive-b-cell-tolerance-by-post-translationally-modified-proteins
#1
Jacqueline S Dekkers, Marije K Verheul, Jeroen N Stoop, Bisheng Liu, Andreea Ioan-Facsinay, Peter A van Veelen, Arnoud H de Ru, George M C Janssen, Martin Hegen, Steve Rapecki, Tom W J Huizinga, Leendert A Trouw, René E M Toes
OBJECTIVES: Over 50% of patients with rheumatoid arthritis (RA) harbour a variety of anti-modified protein antibodies (AMPA) against different post-translationally modified (PTM) proteins, including anti-carbamylated protein (anti-CarP) antibodies. At present, it is unknown how AMPA are generated and how autoreactive B cell responses against PTM proteins are induced. Here we studied whether PTM foreign antigens can breach B cell tolerance towards PTM self-proteins. METHODS: Serum reactivity towards five carbamylated proteins was determined for 160 patients with RA and 40 healthy individuals...
April 25, 2017: Annals of the Rheumatic Diseases
https://www.readbyqxmd.com/read/28407121/urea-and-chronic-kidney-disease-the-comeback-of-the-century-in-uraemia-research
#2
Raymond Vanholder, Tessa Gryp, Griet Glorieux
Urea, a marker of uraemic retention in chronic kidney disease (CKD) and of adequacy of intradialytic solute removal, has traditionally been considered to be biologically inert. However, a number of recent experimental data suggest that urea is toxic at concentrations representative for CKD. First of all, at least five studies indicate that urea itself induces molecular changes related to insulin resistance, free radical production, apoptosis and disruption of the protective intestinal barrier. Second, urea is at the origin of the generation of cyanate, ammonia and carbamylated compounds, which as such all have been linked to biological changes...
April 12, 2017: Nephrology, Dialysis, Transplantation
https://www.readbyqxmd.com/read/28400100/carbamylation-and-oxidation-of-proteins-lead-to-apoptotic-death-of-lymphocytes
#3
Anna Pieniazek, Krzysztof Gwozdzinski
The apoptotic/necrotic changes in isolated human peripheral blood mononuclear cells (MNCs) subjected to hydrogen peroxide (H2O2), cyanate (NaOCN) and their combination were examined. The mitochondrial potential (ΔΨm), the activities of caspases (-2, -3, -6, -8 and -9) and the level of carbonyls and amino groups in proteins were determined and DNA fragmentation. Apoptotic or necrotic cells were identified by fluorescence microscopy using double staining with Hoechst 33258/propidium iodide. Treatment of MNCs with NaOCN (1 mmol/L and 2 mmol/L), alone and in combination with H2O2 (100 μmol/L), led to a significant decrease in the content of amine groups and a significant increase in the carbonyl level of MNCs in comparison with the control...
April 8, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28398552/carbamylated-albumin-is-one-of-the-target-antigens-of-anti-carbamylated-protein-antibodies
#4
Shuichiro Nakabo, Motomu Hashimoto, Shinji Ito, Moritoshi Furu, Hiromu Ito, Takao Fujii, Hajime Yoshifuji, Yoshitaka Imura, Ran Nakashima, Kosaku Murakami, Nobuo Kuramoto, Masao Tanaka, Junko Satoh, Akihito Ishigami, Satoshi Morita, Tsuneyo Mimori, Koichiro Ohmura
Objectives.: Anti-carbamylated protein (anti-CarP) antibodies are detected in RA patients. Fetal calf serum is used as an antigen source in anti-CarP ELISA, and the precise target antigens have not been found. We aimed to identify the target antigens of anti-CarP antibodies. Methods.: Western blotting of anti-CarP antibodies was conducted. Anti-carbamylated human albumin (CarALB) antibody was detected by in-house ELISA for 493 RA patients and 144 healthy controls (HCs)...
April 7, 2017: Rheumatology
https://www.readbyqxmd.com/read/28386136/assessment-of-anti-carp-antibodies-disease-activity-and-quality-of-life-in-rheumatoid-arthritis-patients-on-conventional-and-biological-disease-modifying-antirheumatic-drugs
#5
Sudhir Kumar, Ghanshyam Pangtey, Rachna Gupta, Harmeet Singh Rehan, Lalit Kumar Gupta
OBJECTIVE: Good biomarkers are important to guide decisions in the clinical management of rheumatoid arthritis (RA). RA patients harbor antibodies directed against carbamylated proteins which may predict joint damage. This study investigated whether antibodies against carbamylated proteins (anti-CarP) may serve as surrogate prognostic markers. MATERIAL AND METHODS: Fifty-three patients with a diagnosis of rheumatoid arthritis according to ACR 1987 criteria were included...
2017: Reumatologia
https://www.readbyqxmd.com/read/28382370/impact-of-fibrinogen-carbamylation-on-fibrin-clot-formation-and-stability
#6
Veronika Binder, Brith Bergum, Stéphane Jaisson, Philippe Gillery, Carsten Scavenius, Endy Spriet, Anne Karin Nyhaug, Helen M Roberts, Iain L C Chapple, Annelie Hellvard, Nicolas Delaleu, Piotr Mydel
Carbamylation is a non-enzymatic post-translational modification induced upon exposure of free amino groups to urea-derived cyanate leading to irreversible changes of protein charge, structure and function. Levels of carbamylated proteins increase significantly in chronic kidney disease and carbamylated albumin is considered as an important biomarker indicating mortality risk. High plasma concentrations and long half-life make fibrinogen a prime target for carbamylation. As aggregation and cross-linking of fibrin monomers rely on lysine residues, it is likely that carbamylation impacts fibrinogen processing...
April 6, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28374316/modulation-of-protein-fragmentation-through-carbamylation-of-primary-amines
#7
Sylvester M Greer, Dustin D Holden, Ryan Fellers, Neil L Kelleher, Jennifer S Brodbelt
We evaluate the impact of carbamylation of the primary amines of the side-chains of Lys and the N-termini on the fragmentation of intact protein ions and the chromatographic properties of a mixture of E. coli ribosomal proteins. The fragmentation patterns of the six unmodified and carbamylated proteins obtained by higher energy collision dissociation (HCD) and ultraviolet photodissociation (UVPD) were compared. Carbamylation significantly reduced the total number of protons retained by the protein owing to the conversion of basic primary amines to non-basic carbamates...
April 3, 2017: Journal of the American Society for Mass Spectrometry
https://www.readbyqxmd.com/read/28302542/clinical-implications-associated-with-the-posttranslational-modification-induced-functional-impairment-of-albumin-in-oxidative-stress-related-diseases
#8
REVIEW
Hiroshi Watanabe, Tadashi Imafuku, Masaki Otagiri, Toru Maruyama
Recent research findings indicate that the posttranslational modification of human serum albumin (HSA) such as oxidation, glycation, truncation, dimerization, and carbamylation is related to certain types of diseases. We report herein on a simple and rapid analytical method, using an electrospray ionization time-of-flight mass spectrometry technique, that allows posttranslational modifications of HSA to be quantitatively and qualitatively evaluated with a high degree of sensitivity. In patients with chronic liver disease, chronic renal disease, and diabetes mellitus, an increase in the level of oxidized cysteine-34 (Cys-34) of HSA accompanied by a decrease in the level of reduced Cys-34 was observed...
March 14, 2017: Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28296337/anti-carbamylated-protein-antibodies-in-rheumatoid-arthritis-patients-are-reactive-to-specific-epitopes-of-the-human-fibrinogen-%C3%AE-chain
#9
Jonathan D Jones, B JoNell Hamilton, William F C Rigby
OBJECTIVE: Anti-carbamylated protein antibodies (ACarPA) are associated with rheumatoid arthritis (RA) risk and severity and are primarily directed against fibrinogen. The lack of understanding of ACarPA reactivity has limited analysis of their immunopathogenic associations in RA. To address this shortcoming, we mapped ACarPA epitope reactivity in RA patient sera. METHODS: Immunoblotting identified a patient serum with specific reactivity to carbamylated human fibrinogen β-chain...
March 15, 2017: Arthritis & Rheumatology
https://www.readbyqxmd.com/read/28291659/identification-of-carbamylated-alpha-1-anti-trypsin-a1at-as-an-antigenic-target-of-anti-carp-antibodies-in-patients-with-rheumatoid-arthritis
#10
Marije K Verheul, Alvin Yee, Andrea Seaman, George M Janssen, Peter A van Veelen, Jan W Drijfhout, Rene E M Toes, Michael Mahler, Leendert A Trouw
In 2011 a novel autoantibody system, anti-carbamylated protein (anti-CarP) antibodies, was described in rheumatoid arthritis (RA) patients. Anti-CarP antibody positivity associates with a more severe disease course, is observed years before disease onset, and may predict the development of RA in arthralgia patients. Although many clinical observations have been carried out, information on the antigenic targets of anti-CarP antibodies is limited. Most studies on anti-CarP antibodies utilize an ELISA-based assay with carbamylated fetal calf serum (Ca-FCS) as antigen, a complex mixture of proteins...
March 10, 2017: Journal of Autoimmunity
https://www.readbyqxmd.com/read/28290250/synthesis-and-ativiral-activity-of-5-benzylthio-4-carbamyl-1-2-3-triazoles-against-human-cytomegalovirus-cmv-and-varicella-zoster-virus-vzv
#11
Yi-Ning Wen, Zhi-Feng Zhang, Ning-Ning Liu, Graciela Andrei, Robert Snoeck, Yu-Hong Xiang, Dominique Schols, Xue Chen, Zhuo-Yong Zhang, Qing-Shan Zhang, Qin-Pei Wu
BACKGROUND: All of clinic drugs for herpesvirus infections exhibit high toxicity and suffer from significant drug-resistantance. There is a great need for the development of new, effective, and safe anti-herpesvirus agents with different mechanisms of action. METHODS: A series of novel 5-(benzylthio)-1H-1,2,3- triazole-4-carboxamides were efficiently synthesized and EC50 values against human cytomegalovirus (HCMV), varicella-zoster virus (VZV) and herpes simplex virus (HSV) were evaluated in vitro...
March 7, 2017: Medicinal Chemistry
https://www.readbyqxmd.com/read/28275265/beyond-citrullination-other-post-translational-protein-modifications-in-rheumatoid-arthritis
#12
REVIEW
Leendert A Trouw, Theo Rispens, Rene E M Toes
The presence of autoantibodies is one of the hallmarks of rheumatoid arthritis (RA). In the past few decades, rheumatoid factors (autoantibodies that recognize the Fc-tail of immunoglobulins) as well as anti-citrullinated protein antibodies (ACPAs) have been studied intensively. ACPAs recognize post-translationally modified proteins in which the amino acid arginine has been converted into a citrulline. More recently, other autoantibody systems recognizing post-translationally modified proteins have also gained attention, including autoantibodies recognizing fragmented immunoglobulin (anti-hinge antibodies), autoantibodies recognizing acetylated proteins and autoantibodies recognizing proteins that are modified by adducts formed under oxidative stress...
March 9, 2017: Nature Reviews. Rheumatology
https://www.readbyqxmd.com/read/28183721/carbamylation-of-vimentin-is-inducible-by-smoking-and-represents-an-independent-autoantigen-in-rheumatoid-arthritis
#13
Caroline Ospelt, Holger Bang, Eugen Feist, Giovanni G Camici, Stephan Keller, Jacqueline Detert, Anette Krämer, Steffen Gay, Khetam Ghannam, Gerd R Burmester
OBJECTIVES: Smoking has been connected to citrullination of antigens and formation of anti-citrullinated peptide antibodies (ACPAs) in rheumatoid arthritis (RA). Since smoking can modify proteins by carbamylation (formation of homocitrulline), this study was conducted to investigate these effects on vimentin in animal models and RA. METHODS: The efficiency of enzymatic carbamylation of vimentin was characterised. B-cell response was investigated after immunisation of rabbits with different vimentin isoforms...
February 9, 2017: Annals of the Rheumatic Diseases
https://www.readbyqxmd.com/read/28149015/role-of-bioflavonoid-quercetin-on-expression-of-urea-cycle-enzymes-astrocytic-and-inflammatory-markers-in-hyperammonemic-rats
#14
Sivamani Kanimozhi, Perumal Subramanian, Sakkaravarthy Shanmugapriya, Subramanian Sathishkumar
This study evaluates the role of quercetin on the expression of urea cycle enzymes, astrocytic, neuronal and inflammatory markers in hyperammonemic rats. Hyperammonemia (provoked by intraperitonial injections of (ammonium chloride-100 mg/kg b.w for 56 days), showed diminished expression of urea cycle enzymes [carbamyl phosphate synthetase-1 (CPS-1), ornithine transcarbamylase (OTC), argininosuccinate synthetase (ASS) and arginase (ARG)] in liver and decreased expression of neuronal and astrocytic markers-glutamine synthase (GS) and phosphate activated glutaminase (PAG) in brain and increased expression of brain inflammatory markers such as interleukin 6 (IL6), inducible nitric oxide synthase (iNOS) and nuclear transcription factor kappa B (NF-κB) (by western blot analysis) and exhibited downregulated expression of soluble guanylate cyclase (sGC), glial fibrillary acidic protein (GFAP) in brain and ASS in liver investigated (by RT-PCR)...
March 2017: Indian Journal of Clinical Biochemistry: IJCB
https://www.readbyqxmd.com/read/28137483/differential-effect-of-erythropoietin-and-carbamylated-erythropoietin-on-endothelial-cell-migration
#15
Romina E Maltaneri, María E Chamorro, Agustina Schiappacasse, Alcira B Nesse, Daniela C Vittori
No abstract text is available yet for this article.
January 28, 2017: International Journal of Biochemistry & Cell Biology
https://www.readbyqxmd.com/read/28069651/inhibition-by-avibactam-and-clavulanate-of-the-%C3%AE-lactamases-kpc-2-and-ctx-m-15-harboring-the-substitution-n-132-g-in-the-conserved-sdn-motif
#16
Clément Ourghanlian, Daria Soroka, Michel Arthur
The substitution N(132)G in the SDN motif of class A β-lactamases from rapidly growing mycobacteria was previously shown to impair their inhibition by avibactam but to improve the stability of acyl-enzymes formed with clavulanate. The same substitution was introduced in KPC-2 and CTX-M-15 to assess its impact on β-lactamases from Enterobacteriaceae and evaluate whether it may lead to resistance to the ceftazidime-avibactam combination. Kinetic parameters for the inhibition of the β-lactamases by avibactam and clavulanate were determined by spectrophotometry using nitrocefin as the substrate...
March 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28067524/urea-artifacts-interfere-with-immuno-purification-of-lysine-acetylation
#17
Ana Martinez-Val, Fernando Garcia, Pilar Ximénez-Embún, Ailyn Martínez Teresa-Calleja, Nuria Ibarz, Isabel Ruppen, Javier Munoz
Comprehensive analysis of post-translational modifications (PTMs) often depends on the purification of modified peptides prior to LC-MS/MS. The implementation of these enrichment methods requires thorough knowledge of the experimental conditions to achieve optimal selectivity and sensitivity. In this regard, large-scale analysis of lysine acetylation, a key PTM for multiple cellular processes, makes use of monoclonal pan-antibodies designed against this moiety. We report that the immuno-purification of lysine-acetylated peptides is hampered by the copurification of lysine carbamylated peptides, a frequent urea artifact...
January 17, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28052815/how-do-macrophages-sense-modified-low-density-lipoproteins
#18
REVIEW
Dimitry A Chistiakov, Alexandra A Melnichenko, Alexander N Orekhov, Yuri V Bobryshev
In atherosclerosis, serum lipoproteins undergo various chemical modifications that impair their normal function. Modification of low density lipoprotein (LDL) such as oxidation, glycation, carbamylation, glucooxidation, etc. makes LDL particles more proatherogenic. Macrophages are responsible for clearance of modified LDL to prevent cytotoxicity, tissue injury, inflammation, and metabolic disturbances. They develop an advanced sensing arsenal composed of various pattern recognition receptors (PRRs) capable of recognizing and binding foreign or altered-self targets for further inactivation and degradation...
March 1, 2017: International Journal of Cardiology
https://www.readbyqxmd.com/read/28039278/inhibition-of-%C3%AE-lactamases-of-mycobacteria-by-avibactam-and-clavulanate
#19
Daria Soroka, Clément Ourghanlian, Fabrice Compain, Marion Fichini, Vincent Dubée, Jean-Luc Mainardi, Jean-Emmanuel Hugonnet, Michel Arthur
Objectives: Mycobacterium tuberculosis and Mycobacterium abscessus produce broad-spectrum class A β-lactamases, BlaC and Bla Mab , which are inhibited by clavulanate and avibactam, respectively. BlaC differs from Bla Mab at Ambler position 132 in the conserved motif SDN (SDG versus SDN, respectively). Here, we investigated whether this polymorphism could account for the inhibition specificity of β-lactamases from slowly and rapidly growing mycobacteria. Methods: Enzyme kinetics were determined to assess the impact of the substitutions G 132 N in BlaC and N 132 G in Bla Mab on β-lactamase inhibition by clavulanate and avibactam...
April 1, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28007335/precision-medicine-in-rare-disease-mechanisms-of-disparate-effects-of-n-carbamyl-l-glutamate-on-mutant-cps1-enzymes
#20
Dashuang Shi, Gengxiang Zhao, Nicholas Ah Mew, Mendel Tuchman
This study documents the disparate therapeutic effect of N-carbamyl-l-glutamate (NCG) in the activation of two different disease-causing mutants of carbamyl phosphate synthetase 1 (CPS1). We investigated the effects of NCG on purified recombinant wild-type (WT) mouse CPS1 and its human corresponding E1034G (increased ureagenesis on NCG) and M792I (decreased ureagenesis on NCG) mutants. NCG activates WT CPS1 sub-optimally compared to NAG. Similar to NAG, NCG, in combination with MgATP, stabilizes the enzyme, but competes with NAG binding to the enzyme...
March 2017: Molecular Genetics and Metabolism
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