Related ArticlesDetermination of NADH-dependent glutamate synthase (GOGAT) in Spodoptera frugiperda (Sf9) insect cells by a selective 1H/15N NMR in vitro assay.
J Biotechnol. 2000 Apr 14;79(1):87-97
Authors: Doverskog M, Jacobsson U, Chapman BE, Kuchel PW, Häggström L
This is the second of two papers [Drews, M., Doverskog, M., Ohman, L., Chapman, B.E., Jacobsson, U., Kuchel, P.W., Häggström, L., 2000. Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR. J. Biotechnol. 78, 23-37]. where the general goal has been to determine and characterise the glutamine metabolism in Sf9 cells. The presence of glutamate synthase (GOGAT) activity was investigated in cell-free extracts of S. frugiperda (Sf9) insect cells by modified 1H/15N spin-echo and gradient enhanced multiple quantum coherence NMR spectroscopy techniques. Cell-free extracts were prepared from cells cultured in a serum-free medium. The assay conditions were based on conventional spectrophotometric and chromatographic methods. NMR data showed that nitrogen from [5-15N] glutamine was selectively incorporated into 2-oxoglutarate forming [2-15N] glutamate with a specific activity of 4.15 +/- 0.21 nmol [2-15N] glutamate min -1 (mg total protein)-1 in the cell-free extracts. The enzyme activity was exclusively dependent on NADH as coenzyme and was completely inhibited by 1 mM azaserine. From the results obtained, we conclude that Sf9 cells possess NADH-GOGAT activity. Furthermore, the high specificity of the NMR method enables distinction of competing reactions from glutaminase and glutamate dehydrogenase.
Active site dynamics in NADH oxidase from Thermus thermophilus studied by NMR spin relaxation
Active site dynamics in NADH oxidase from Thermus thermophilus studied by NMR spin relaxation
Abstract We have characterized the backbone dynamics of NADH oxidase from Thermus thermophilus (NOX) using a recently-developed suite of NMR experiments designed to isolate exchange broadening, together with 15N R 1, R 1Ï? , and {1H}-15N steady-state NOE relaxation measurements performed at 11.7 and 18.8 T. NOX is a 54 kDa homodimeric enzyme that belongs to a family of structurally homologous flavin reductases and nitroreductases with many potential biotechnology applications. Prior studies...
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Role of aminotransferases in glutamate metabolism of human erythrocytes
Role of aminotransferases in glutamate metabolism of human erythrocytes
Abstract Human erythrocytes require a continual supply of glutamate to support glutathione synthesis, but are unable to transport this amino acid across their cell membrane. Consequently, erythrocytes rely on de novo glutamate biosynthesis from α-ketoglutarate and glutamine to maintain intracellular levels of glutamate. Erythrocytic glutamate biosynthesis is catalyzed by three enzymes, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and glutamine aminohydrolase (GA). Although the presence of these...
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NMR assignment of 1H, 13C, and 15N resonances of rat lipocalin-type prostaglandin D synthase.
NMR assignment of 1H, 13C, and 15N resonances of rat lipocalin-type prostaglandin D synthase.
NMR assignment of 1H, 13C, and 15N resonances of rat lipocalin-type prostaglandin D synthase.
Biomol NMR Assign. 2010 Oct;4(2):223-5
Authors: Liu J, Lv Y, Guo C, Lin D
Lipocalin-type prostaglandin D synthase (L-PGDS) acts as both a PGD(2)-synthesizing enzyme and an extracellular transporter for small lipophilic molecules. Here we report the backbone and side-chain resonance assignments of uniformly (15)N, (13)C labeled rat L-PGDS.
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[NMR paper] Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha.
Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha.
Related Articles Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha.
Protein Sci. 2005 Sep;14(9):2421-8
Authors: Schnell JR, Zhou GP, Zweckstetter M, Rigby AC, Chou JJ
Coiled-coil motifs play essential roles in protein assembly and molecular recognition, and are therefore the targets of many ongoing...
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12-01-2010 06:56 PM
[NMR paper] Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with pr
Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent.
Related Articles Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent.
FEBS Lett. 2004 Jan 2;556(1-3):35-8
Authors: Dmitriev OY, Altendorf K, Fillingame RH
Subunit a of the Escherichia coli ATP synthase, a 30 kDa integral membrane protein, was purified to homogeneity by a novel procedure incorporating selective extraction into a monophasic...
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[NMR paper] Measurement of side-chain carboxyl pK(a) values of glutamate and aspartate residues i
Measurement of side-chain carboxyl pK(a) values of glutamate and aspartate residues in an unfolded protein by multinuclear NMR spectroscopy.
Related Articles Measurement of side-chain carboxyl pK(a) values of glutamate and aspartate residues in an unfolded protein by multinuclear NMR spectroscopy.
J Am Chem Soc. 2002 May 22;124(20):5714-7
Authors: Tollinger M, Forman-Kay JD, Kay LE
A triple-resonance NMR pulse scheme is presented for measuring aspartic and glutamic acid side-chain pK(a) values in unfolded protein states where chemical shift...
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[NMR paper] 13C NMR analysis of the cysteine-sulfenic acid redox center of enterococcal NADH pero
13C NMR analysis of the cysteine-sulfenic acid redox center of enterococcal NADH peroxidase.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles 13C NMR analysis of the cysteine-sulfenic acid redox center of enterococcal NADH peroxidase.
Biochemistry. 1997 Jul 15;36(28):8611-8
Authors: Crane EJ, Vervoort J, Claiborne A
In order to characterize the native Cys42-sulfenic acid redox center of the flavoprotein NADH peroxidase by NMR, an expression protocol has been developed which yields the...
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[NMR paper] 1H- and 13C-NMR investigation of redox-state-dependent and temperature-dependent conf
1H- and 13C-NMR investigation of redox-state-dependent and temperature-dependent conformation changes in horse cytochrome c.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles 1H- and 13C-NMR investigation of redox-state-dependent and temperature-dependent conformation changes in horse cytochrome c.
Eur J Biochem. 1993 Feb 1;211(3):555-62
Authors: Turner DL, Williams RJ
The redox-state dependent changes in chemical shift, which have...
Determination of NADH-dependent glutamate synthase (GOGAT) in Spodoptera frugiperda (
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