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2 edition of Structural and functional studies of folylpolyglutamate synthetase. found in the catalog.

Structural and functional studies of folylpolyglutamate synthetase.

Yi Sheng

Structural and functional studies of folylpolyglutamate synthetase.

by Yi Sheng

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Published .
Written in English


About the Edition

Folylpolyglutamate synthetase (FPGS) catalyzes the polyglutamylation of folates and antifolates. FPGS is an essential enzyme, required in bacteria for folate biosynthesis and in mammals for folate retention in cells and in organelles, and itself is also a potential drug target for cancer chemotherapy and treatment of microbial infection. I have undertaken structural and functional studies of FPGS with the goal of gaining better understanding of the mechanism of FPGS.Using Electronic Paramagnetic Resonance (EPR) and the site-directed spin labeling (SDSL) technique, I detected conformational changes of FPGS upon binding ATP or folate. This study not only complements the crystal structure data but also provided dynamic information on FPGS in the solution state for the first time.I have characterized the conserved residues at the active site of Lactobacillus casei (L. casei) FPGS, using site-directed mutagenesis, enzyme assays and kinetic experiments. This study provided biochemical information for the folate- and glutamate-binding sites of FPGS and determined the functional roles for each mutated residue.The difference between the L. casei FPGS and E. coli enzyme lies in the fact that the former has only FPGS activity and the latter functions as both FPGS and dihydrofolate synthetase. I have made chimeric proteins of L. casei and E. coli FPGS and characterized them using enzyme specific activity assay, equilibrium dialysis and fluorescence. This study showed that there is no additional domain required for DHFS activity. In addition, it provided information on the structural determinants for folate substrate specificity.I have characterized an essential glutamate residue of FPGS (Glu143 in L. casei FPGS), which has been proposed to coordinate the Mg 2+ ion in the crystal structure of L. casei FPGS. Using site-directed mutagenesis, equilibrium dialysis and Thin-layer chromatography (TLC) assay, I was able to show that the glutamate mutant had increased ATPase activity and was defective in transferring the gamma-phosphate of ATP to folate. This result not only demonstrated the importance of Glu143 in FPGS function, but also shed light on the working mechanism of enzyme catalysis.

The Physical Object
Pagination209 leaves.
Number of Pages209
ID Numbers
Open LibraryOL22609976M
ISBN 100612917657

Folylpolyglutamate synthetase (FPGS) catalyzes the addition of glutamate residues to folates and antifolates to form the physiological active coenzymatic forms of the vitamin and more potent anti-folate agents (reviewed in reference 1). Studies on FPGS have been hampered by the low abundance and instability of the by: 5.   The structure of the apo thiolation-thioesterase di-domain fragment of the EntF non-ribosomal peptide synthetase subunit of enterobactin synthetase is solved. Extensive inter- and intra-domain Cited by:

  The mechanism of sensing and signalling of cytosolic DNA by the innate immune system is a topic of intense research interest as it is the means by which invading bacteria and viruses are by:   INTRODUCTION. The enzyme folylpolyglutamate synthetase (FPGS) 1 in mammalian cells has both physiological and pharmacological significance. FPGS mediates the anabolism of folates and their analogs to γ-polyglutamate peptides (1–5).Polyglutamated forms of folate coenzymes are conserved and are more efficient as cofactors for folate-dependent .

S-Adenosylmethionine synthetase (ATP: l-methionine S-adenosyltransferase) catalyzes a two-step reaction in which tripolyphosphate (PPPi) is a tightly bound intermediate. Diimidotriphosphate (O3P-NH-PO2-NH-PO3; PNPNP), a non-hydrolyzable analogue of PPPi, is the most potent known inhibitor of AdoMet synthetase with a Ki of 2 nM. The structural basis for the slow, .   Tetrahydrofolate coenzymes involved in one-carbon (C1) metabolism are polyglutamylated. In organisms that synthesize tetrahydrofolate de novo, dihydrofolate synthetase (DHFS) and folylpolyglutamate synthetase (FPGS) catalyze the attachment of glutamate residues to the folate this study we isolated cDNAs coding a DHFS and Cited by:


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Structural and functional studies of folylpolyglutamate synthetase by Yi Sheng Download PDF EPUB FB2

Structural and functional similarities in the ADP-forming amide bond ligase superfamily: implications for a substrate-induced conformational change in folylpolyglutamate synthetase 1 1Edited by I. Wilson. Journal of Molecular Biology(2), DOI: /jmbiCited by: The Folylpolyglutamate synthetase with ligands was placed into a simulation box with appropriate buffer, or other solutions, at a minimum distance of Å from the complex.

The solution for simulation was based on the TIP3P water model in which sodium and chloride ions were added to neutralize complex by: 1.

Sheng Y, Sun X, Shen Y, Bognar AL, Baker EN, Smith CA. Structural and functional similarities in the ADP-forming amide bond ligase superfamily: implications for a substrate-induced conformational change in folylpolyglutamate synthetase.

J Mol Biol –, PubMed CrossRef Google ScholarAuthor: Yi Sheng, Yang Shen, Andrew L. Bognar. In Neurospora crassa, the met-6 + gene encodes folylpoly-γ-glutamate synthetase (FPGS) which catalyzes the formation of polyglutamate forms of nine auxotrophy of the Neurospora crassa met-6 mutant is related to a lesion affecting this enzyme.

Functional complementation of the mutant strain was achieved by introducing copies of the wild-type met-6 + gene into Cited by: 7. @article{osti_, title = {Loss of function of folylpolyglutamate synthetase 1 reduces lignin content and improves cell wall digestibility in Arabidopsis}, author = {Srivastava, Avinash C.

and Chen, Fang and Ray, Tui and Pattathil, Sivakumar and Peña, Maria J. and Avci, Utku and Li, Hongjia and Huhman, David V. and Backe, Jason and.

The three-dimensional structure of folylpolyglutamate synthase of Lactobacillus casei and E. coli have been determined by X-ray crystallography.

The catalytically active conformation of the protein is obtained by bonding of a folate-type substrate; binding of ATP is not, per se, sufficient to generate that conformational state. The folylpolyglutamate synthetase-dihydrofolate synthetase gene (folC) in Escherichia coli was deleted from the bacterial chromosome and replaced by a selectable Kmr marker.

The deletion strain required a complementing gene expressing folylpolyglutamate synthetase encoded on a plasmid for viability, indicating that folC is an essential gene in Cited by: Folylpolyglutamate synthase, mitochondrial is an enzyme that in humans is encoded by the FPGS gene.

This gene encodes the folylpolyglutamate synthetase enzyme. This enzyme has a central role in establishing and maintaining both cytosolic and mitochondrial folylpolyglutamate concentrations and, therefore, is essential for folate homeostasis and the survival of Aliases: FPGS, folylpolyglutamate synthase.

Functional studies on FAALs from Mtb (MtFAAL) documented that the enzymes activate fatty acids as acyl-adenylates, but do not catalyze thioester-forming reactions 5, 6.

It was therefore suggested that the acyl-adenylates serve as substrates for multifunctional PKSs to permit synthesis of complex lipids such as phthiocerol dimycocerosate (PDIM Cited by: Comparison of the three-dimensional structures of folylpolyglutamate synthetase (FPGS) and the bacterial cell wall ligase UDP-N-acetylmuramoyl-l-alanine: d-glutamate ligase (MurD) reveals that these two enzymes have a remarkable structural similarity despite a low level of sequence enzymes have a modular, multi-domain structure and catalyse a similar ATP Cited by:   Structural Studies of Lysyl-tRNA Synthetase: Conformational Changes Induced by Substrate Binding.

Biochemistry39 (42), DOI: /bir. Dénes Berta, Pedro J Buigues, Magd Badaoui, Edina by:   Drake EJ, Nicolai DA, Gulick AM () Structure of the EntB multidomain nonribosomal peptide synthetase and functional analysis of its interaction with the EntE adenylation domain.

Chem Biol – CrossRef PubMed Google ScholarCited by:   Folylpolyglutamate synthase is a major determinant of intracellular methotrexate polyglutamates in patients with rheumatoid arthritis.

Sci. Rep. 6, Cited by: FolC plays important roles in the folate metabolism of cells by attaching l-Glu to dihydropteroate (DHP) and folate, which are known activities of dihydrofolate synthetase (DHFS) and folylpolyglutamate synthetase (FPGS), respectively. Here, we determined the crystal structure of Helicobacter pylori FolC (HpFolC) at Å resolution using the single-wavelength Cited by: 1.

Structural Studies on Formyl-Methenyl-Methylene Tetrahydrofolate Synthetase from Rabbit Liver; Utilization of Folate Polyglutamate for Growth by Lactobacillus Casei; Folate and Folate Enzymes as Structural Components of Bacteriophage Particles; Studies on the Substrate Specificity of Mammalian Folylpolyglutamate Synthetase.

Folate polyglutamylation is carried out by the enzyme folylpolyglutamate synthetase (FPGS). Initial gene cloning, functional characterization, and cellular localization of the three isoforms of FPGS [FPGS1 (plastid), FPGS2 (mitochondria), and FPGS3 (cytosol)] in Arabidopsis were described by Ravanel et by: Aminoacyl-tRNA synthetases (AARSs) are the enzymes that catalyze the aminoacylation reaction by covalently linking an amino acid to its cognate tRNA in the first step of protein translation.

Beyond this classical function, these enzymes are also known to have a role in several metabolic and signaling pathways that are important for cell by: Pergamon Press pic STRUCTURAL SPECIFICITY OF INHIBITION OF HUMAN FOLYLPOLYGLUTAMATE SYNTHETASE BY ORNITHINE- CONTAINING FOLATE ANALOGS JOHN J.

McGuiRE,*f WANDA E. BOLANOWSKA* and JAMES R. PIPERÎ * Grace Cancer Drug Center, Roswell Park Memorial Institute, Buffalo, NY ; and t Drug Synthesis Branch, Cited by: The Inhibition of Folylpolyglutamate Synthetase (folC) in the Prevention of Drug Resistance in Mycobacterium tuberculosis by Traditional Chinese Available.

Abstract. CCRF-CEM human leukemia cell lines resistant to “intermittent”, but not continuous, exposure to methotrexate (MTX) as a result of defective MTX polyglutamate synthesis were recently described defective synthesis was traced to decreased folylpolyglutamate synthetase (FPGS) activity; the FPGS level was inversely related to the degree of resistance by: 4.

Studies on the Substrate Specificity of Mammalian Folylpolyglutamate Synthetase McGuire, J. J. / Hsieh, P. / Bertino, J. R. / Coward, J. Κ.In enzymology, a tetrahydrofolate synthase (EC ) Structural studies. As of latemolecular and functional characterization of dihydrofolate synthetase and three isoforms of folylpolyglutamate synthetase in Arabidopsis thaliana".

Proc. Natl. Acad. : BRENDA entry.For this special issue, “Crystallographic Studies of Enzymes", we have collected research papers on enzymes with structural aspects and functional aspects; here we briefly discuss the contents of such research papers as follows, with the aim of suggesting new directions of investigation in the fields of enzyme research, protein engineering.