Design of an in vitro multienzyme cascade system for the biosynthesis of nicotinamide mononucleotide

2022 ◽  
Author(s):  
Cailian Zhou ◽  
Jiao Feng ◽  
Jing Wang ◽  
Ning Hao ◽  
Xin Wang ◽  
...  
Keyword(s):  
Cascade System ◽  
Nicotinamide Mononucleotide ◽  
Phosphate Hydrolysis ◽  
Adenosine Phosphate

Design the adenosine phosphate hydrolysis (APH) pathway multienzyme cascade system for the biosynthesis of nicotinamide mononucleotide (NMN) in vitro.

scholarly journals Detection and pharmacological modulation of nicotinamide mononucleotide (NMN) in vitro and in vivo

2009 ◽  
Vol 77 (10) ◽  
pp. 1612-1620 ◽  
Author(s):  
Laura Formentini ◽  
Flavio Moroni ◽  
Alberto Chiarugi
Keyword(s):  
Pharmacological Modulation ◽  
Nicotinamide Mononucleotide

The cascade catalysis of the porphyrinic zirconium metal-organic framework PCN-224-Cu for CO2 conversion to alcohols

2021 ◽  
Author(s):  
Shi-Chao Qi ◽  
Zhi-Hui Yang ◽  
Rong-Rong Zhu ◽  
Xiao-Jie Lu ◽  
Ding-Ming Xue ◽  
...  
Keyword(s):  
Metal Organic Framework ◽  
Co2 Conversion ◽  
Cascade System ◽  
The Past ◽  
Highly Efficient ◽  
Metal Organic ◽  
Zirconium Metal ◽  
Organic Framework ◽  
Cascade Catalysis

Reproducing the highly efficient catalytic cascade system to mimic life behaviors in vitro has been intensively investigated over the past few decades. However, except the biocatalysis with enzymes, most chemocatalytic...

scholarly journals Affinity chromatography of nicotinamide nucleotide-dependent dehydrogenases on immobilized nucleotide derivatives

1974 ◽  
Vol 141 (3) ◽  
pp. 775-787 ◽  
Author(s):  
Ian P. Trayer ◽  
Hylary R. Trayer
Keyword(s):  
Affinity Chromatography ◽  
Glucose Dehydrogenase ◽  
Three Dimensional ◽  
Ligand Affinity ◽  
Nicotinamide Mononucleotide ◽  
Adenosine Phosphate ◽  
Three Dimensional Models ◽  
Separation Of Mixtures ◽  
Phosphate Ligands ◽  
Sepharose 4B

A series of chemically-defined adenosine phosphate ligands attached to Sepharose 4B were used as active-site probes in studying the interaction of enzymes with their coenzymes and substrates and to test the suitability of these matrices for ‘general ligand’ affinity chromatography. Nicotinamide nucleotide-dependent dehydrogenases were used as models to test this methodology. Elution from these columns by NAD+and/or AMP gradients (in the presence or the absence of substrates and/or nicotinamide mononucleotide) was consistent with: (1) the compulsory ordered addition of substrates to lactate and malate dehydrogenase; (2) the necessity for the NMN moiety of NAD+to bind to these enzymes before the substrate; and illustrated: (3) that the binding of these two hydrogenases to these columns compared very well with the published three-dimensional models for these enzymes and (4) that separation of mixtures of dehydrogenases depended on the choice of matrix and displacing ion and whether any additions (e.g. substrates) were made to the gradients used. These techniques were used to purify UDP-glucose dehydrogenase from a crude starting material on a phosphate-linked UDP (or ADP) matrix. The binding of this enzyme to these two columns was not consistent with either an ordered or random addition of substrates and suggested a more complex mechanism.

scholarly journals Comparative Genomics of NAD Biosynthesis in Cyanobacteria

2006 ◽  
Vol 188 (8) ◽  
pp. 3012-3023 ◽  
Author(s):  
Svetlana Y. Gerdes ◽  
Oleg V. Kurnasov ◽  
Konstantin Shatalin ◽  
Boris Polanuyer ◽  
Roman Sloutsky ◽  
...  
Keyword(s):  
De Novo ◽  
Physiological Role ◽  
Model Organisms ◽  
Special Importance ◽  
Nicotinamide Mononucleotide ◽  
Genomic Platform ◽  
Salvage Pathways ◽  
Conserved Gene ◽  
Key Aspects

ABSTRACT Biosynthesis of NAD(P) cofactors is of special importance for cyanobacteria due to their role in photosynthesis and respiration. Despite significant progress in understanding NAD(P) biosynthetic machinery in some model organisms, relatively little is known about its implementation in cyanobacteria. We addressed this problem by a combination of comparative genome analysis with verification experiments in the model system of Synechocystis sp. strain PCC 6803. A detailed reconstruction of the NAD(P) metabolic subsystem using the SEED genomic platform (http://theseed.uchicago.edu/FIG/index.cgi ) helped us accurately annotate respective genes in the entire set of 13 cyanobacterial species with completely sequenced genomes available at the time. Comparative analysis of operational variants implemented in this divergent group allowed us to elucidate both conserved (de novo and universal pathways) and variable (recycling and salvage pathways) aspects of this subsystem. Focused genetic and biochemical experiments confirmed several conjectures about the key aspects of this subsystem. (i) The product of the slr1691 gene, a homolog of Escherichia coli gene nadE containing an additional nitrilase-like N-terminal domain, is a NAD synthetase capable of utilizing glutamine as an amide donor in vitro. (ii) The product of the sll1916 gene, a homolog of E. coli gene nadD, is a nicotinic acid mononucleotide-preferring adenylyltransferase. This gene is essential for survival and cannot be compensated for by an alternative nicotinamide mononucleotide (NMN)-preferring adenylyltransferase (slr0787 gene). (iii) The product of the slr0788 gene is a nicotinamide-preferring phosphoribosyltransferase involved in the first step of the two-step nondeamidating utilization of nicotinamide (NMN shunt). (iv) The physiological role of this pathway encoded by a conserved gene cluster, slr0787-slr0788, is likely in the recycling of endogenously generated nicotinamide, as supported by the inability of this organism to utilize exogenously provided niacin. Positional clustering and the cooccurrence profile of the respective genes across a diverse collection of cellular organisms provide evidence of horizontal transfer events in the evolutionary history of this pathway.

scholarly journals Mn(III) Pyrophosphate as an Efficient Tool for Studying the Mode of Action of Isoniazid on the InhA Protein of Mycobacterium tuberculosis

2002 ◽  
Vol 46 (7) ◽  
pp. 2137-2144 ◽  
Author(s):  
Michel Nguyen ◽  
Annaïk Quémard ◽  
Sylvain Broussy ◽  
Jean Bernadou ◽  
Bernard Meunier
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Biosynthetic Pathway ◽  
Acyl Carrier Protein ◽  
Isonicotinic Acid ◽  
Structural Investigations ◽  
Mycolic Acids ◽  
Nicotinamide Mononucleotide ◽  
Catalase Peroxidase ◽  
Nicotinamide Coenzymes

ABSTRACT The antituberculosis drug isoniazid (INH) is quickly oxidized by stoichiometric amounts of manganese(III) pyrophosphate. In the presence of nicotinamide coenzymes (NAD+, NADH, nicotinamide mononucleotide [NMN+]) and nicotinic acid adenine dinucleotide (DNAD+), INH oxidation produced the formation of INH-coenzyme adducts in addition to known biologically inactive products (isonicotinic acid, isonicotinamide, and isonicotinaldehyde). A pool of INH-NAD(H) adducts preformed in solution allowed the rapid and strong inhibition of in vitro activity of the enoyl-acyl carrier protein reductase InhA, an INH target in the biosynthetic pathway of mycolic acids: the inhibition was 90 or 60% when the adducts were formed in the presence of NAD+ or NADH, respectively. Under similar conditions, no inhibitory activity of INH-NMN(H) and INH-DNAD(H) adducts was detected. When an isolated pool of 100 nM INH-NAD(H) adducts was first incubated with InhA, the enzyme activity was inhibited by 80%; when present in excess, both NADH and decenoyl-coenzyme A are able to prevent this phenomenon. InhA inhibition by several types of INH-coenzyme adducts coexisting in solution is discussed in relation with the structure of the coenzyme, the stereochemistry of the adducts, and their existence as both open and cyclic forms. Thus, manganese(III) pyrophosphate appears to be an efficient and convenient alternative oxidant to mimic the activity of the Mycobacterium tuberculosis KatG catalase-peroxidase and will be useful for further mechanistic studies of INH activation and for structural investigations of reactive INH species in order to promote the design of new inhibitors of InhA as potential antituberculous drugs.

scholarly journals A Novel NAD-RNA Decapping Pathway Discovered by Synthetic Light-Up NAD-RNAs

Biomolecules ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 513
Author(s):  
Florian Abele ◽  
Katharina Höfer ◽  
Patrick Bernhard ◽  
Julia Grawenhoff ◽  
Maximilian Seidel ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Rna Degradation ◽  
Enzymatic Reactions ◽  
Nicotinamide Mononucleotide ◽  
Decapping Enzyme ◽  
New Research ◽  
Rna Decapping ◽  
Decapping Enzymes

The complexity of the transcriptome is governed by the intricate interplay of transcription, RNA processing, translocation, and decay. In eukaryotes, the removal of the 5’-RNA cap is essential for the initiation of RNA degradation. In addition to the canonical 5’-N7-methyl guanosine cap in eukaryotes, the ubiquitous redox cofactor nicotinamide adenine dinucleotide (NAD) was identified as a new 5’-RNA cap structure in prokaryotic and eukaryotic organisms. So far, two classes of NAD-RNA decapping enzymes have been identified, namely Nudix enzymes that liberate nicotinamide mononucleotide (NMN) and DXO-enzymes that remove the entire NAD cap. Herein, we introduce 8-(furan-2-yl)-substituted NAD-capped-RNA (FurNAD-RNA) as a new research tool for the identification and characterization of novel NAD-RNA decapping enzymes. These compounds are found to be suitable for various enzymatic reactions that result in the release of a fluorescence quencher, either nicotinamide (NAM) or nicotinamide mononucleotide (NMN), from the RNA which causes a fluorescence turn-on. FurNAD-RNAs allow for real-time quantification of decapping activity, parallelization, high-throughput screening and identification of novel decapping enzymes in vitro. Using FurNAD-RNAs, we discovered that the eukaryotic glycohydrolase CD38 processes NAD-capped RNA in vitro into ADP-ribose-modified-RNA and nicotinamide and therefore might act as a decapping enzyme in vivo. The existence of multiple pathways suggests that the decapping of NAD-RNA is an important and regulated process in eukaryotes.

scholarly journals Nicotinamide mononucleotide adenylyltransferase uses its NAD+ substrate-binding site to chaperone phosphorylated Tau

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Xiaojuan Ma ◽  
Yi Zhu ◽  
Jinxia Lu ◽  
Jingfei Xie ◽  
Chong Li ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Single Molecule ◽  
Molecular Chaperones ◽  
Phosphorylated Tau ◽  
Amyloid Aggregation ◽  
Computational Approaches ◽  
Nad Metabolism ◽  
Nicotinamide Mononucleotide ◽  
Nicotinamide Mononucleotide Adenylyltransferase

Tau hyper-phosphorylation and deposition into neurofibrillary tangles have been found in brains of patients with Alzheimer’s disease (AD) and other tauopathies. Molecular chaperones are involved in regulating the pathological aggregation of phosphorylated Tau (pTau) and modulating disease progression. Here, we report that nicotinamide mononucleotide adenylyltransferase (NMNAT), a well-known NAD+ synthase, serves as a chaperone of pTau to prevent its amyloid aggregation in vitro as well as mitigate its pathology in a fly tauopathy model. By combining NMR spectroscopy, crystallography, single-molecule and computational approaches, we revealed that NMNAT adopts its enzymatic pocket to specifically bind the phosphorylated sites of pTau, which can be competitively disrupted by the enzymatic substrates of NMNAT. Moreover, we found that NMNAT serves as a co-chaperone of Hsp90 for the specific recognition of pTau over Tau. Our work uncovers a dedicated chaperone of pTau and suggests NMNAT as a key node between NAD+ metabolism and Tau homeostasis in aging and neurodegeneration.

scholarly journals Mechanistic insights into NAD synthase NMNAT chaperoning phosphorylated Tau from pathological aggregation

2019 ◽  
Author(s):  
Xiaojuan Ma ◽  
Jinxia Lu ◽  
Yi Zhu ◽  
Jingfei Xie ◽  
Chong Li ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Single Molecule ◽  
Neurofibrillary Tangles ◽  
Molecular Chaperones ◽  
Phosphorylated Tau ◽  
Amyloid Aggregation ◽  
Computational Approaches ◽  
Nad Metabolism ◽  
Nicotinamide Mononucleotide

AbstractTau hyper-phosphorylation and deposition into neurofibrillary tangles have been found in brains of patients with Alzheimer’s disease (AD) and other tauopathies. Molecular chaperones are involved in regulating the pathological aggregation of phosphorylated Tau (pTau) and modulating disease progression. Here, we report that nicotinamide mononucleotide adenylyltransferase (NMNAT), a well-known NAD synthase, serves as a chaperone of pTau to prevent its amyloid aggregation in vitro as well as mitigate its pathology in a fly tauopathy model. By combining NMR spectroscopy, crystallography, single-molecule and computational approaches, we revealed that NMNAT adopts its enzymatic pocket to specifically bind the phosphorylated sites of pTau, which can be competitively disrupted by the enzymatic substrates of NMNAT. Moreover, we found that NMNAT serves as a co-chaperone of Hsp90 for the specific recognition of pTau over Tau. Our work uncovers a dedicated chaperone of pTau and suggests NMNAT as a key node between NAD metabolism and Tau homeostasis in aging and neurodegeneration.

scholarly journals Anti-oxidative effects of nicotinamide mononucleotide, a regulator of aging, on rat high glucose-induced tenocytes in vitro

2020 ◽  
Author(s):  
Kohei Yamaura ◽  
Yutaka Mifune ◽  
Atsuyuki Inui ◽  
Hanako Nishimoto ◽  
Takeshi Kataoka ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
High Glucose ◽  
Messenger Rna ◽  
Oxygen Species ◽  
Sprague Dawley ◽  
Stress Effects ◽  
Nicotinamide Mononucleotide ◽  
Sprague Dawley Rats ◽  
Oxidative Effects

Abstract Background Nicotinamide adenine dinucleotide (NAD+) plays an important role in energy metabolism, mitochondrial function, aging, and cell death. Nicotinamide mononucleotide (NMN) is one of the key precursors of NAD+. The purpose of this study is to evaluate the oxidative stress effects of NMN on rat tenocytes in-vitro.Methods Tenocytes from normal Sprague–Dawley rats were cultured in regular glucose (RG) and high-glucose (HG) conditions with or without NMN, and were divided into four groups: RG NMN−, RG NMN+, HG NMN−, and HG NMN+. Cell viability, reactive oxygen species (ROS) production, apoptosis, and messenger RNA (mRNA) expressions of NADPH oxidase (NOX) 1, NOX4, interleukin (IL)-6, SIRT1, and SIRT6, were determined in-vitro.Results The NMN groups led to significantly higher cell viabilities compared with the other groups. The mRNA expressions of NOX1, NOX4, and IL6, in the HG NMN+ group were significantly lower compared with those of the HG NMN− group. Conversely, the corresponding expressions of the SIRT1 and SIRT6 levels in the HG NMN+ group were significantly higher compared with those of the HG NMN−group. Both the accumulation of ROS and apoptosis in the HG NMN− group were significantly higher compared with those in the RG NMN− group at 48 h.Conclusion The expression levels of NOX1, NOX4, IL6, and ROS were significantly reduced by NMN. These results suggest that NMN could effectively reduce the oxidative stress by activating SIRT1 and SIRT6, and by inhibiting the activity of NOX and apoptosis in the tenocytes.

Sign in / Sign up

Export Citation Format

Share Document