Column chromatography of amino acids with fluorescence detection

Roth, Marc; Hampai, A

doi:10.1016/s0021-9673(00)97051-1

Cited by 295 publications

(63 citation statements)

References 2 publications

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“…At 34 mo after OLT, amino acids were determined by column chromatography (7). Amino acid separation was performed by a method modified from Hamilton (8).…”

Section: Amino Acidsmentioning

confidence: 99%

Orthotopic liver transplantation for urea cycle enzyme deficiency

et al. 1992

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Hyperammonemia, abnormalities in plasma amino acids and abnormalities of standard liver functions were corrected by orthotopic liver transplantation in a 14-day-old boy with carbamyl phosphate synthetase-I deficiency and in a 35-yr-old man with argininosuccinic acid synthetase deficiency. The first patient had high plasma glutamine levels and no measureable citrulline, whereas citrulline values were markedly increased in Patient 2. Enzyme analysis of the original livers showed undetectable activity of carbamyl phosphate synthetase-I in Patient 1 and arginosuccinic acid synthetase in Patient 2. Both patients were comatose before surgery. Intellectual recovery of patient 1 has been slightly retarded because of a brain abscess caused by Aspergillus infection after surgery. Both patients are well at 34 and 40 mo, respectively, after surgery. Our experience has shown that orthotopic liver transplantation corrects the life-threatening metabolic abnormalities caused by deficiencies in the urea cycle enzymes carbamyl phosphate synthetase-I and arginosuccinic acid synthetase. Seven other patients-six with ornithine transcarbamylase deficiency and another with carbamyl phosphate synthetase-I deficiency-are known to have been treated elsewhere with liver transplantation 1½ yr or longer ago. Four of these seven recipients also are well, with follow-ups of 1½ to 5 yr. Thus liver transplantation corrects the metabolic abnormalities of three of the six urea cycle enzyme deficiencies, and presumably would correct all.

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“…At 34 mo after OLT, amino acids were determined by column chromatography (7). Amino acid separation was performed by a method modified from Hamilton (8).…”

Section: Amino Acidsmentioning

confidence: 99%

Orthotopic liver transplantation for urea cycle enzyme deficiency

et al. 1992

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“…The analyses were done with either ninhydrin methodology (Beckman 119 automatic analyser) or using hypochlorite and o-phthalaldehyde post-column fluorescence detection (Waters Associates HPLC amino acid analyser [9]). …”

Section: Analytical Proceduresmentioning

confidence: 99%

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

A¹,

Wf²,

C³

1985

Eur J Biochem

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1. The complete amino acid sequence (145 residues) of histone variant H2A,,, from wheat germ Triticum 2. The sequence of histone variant H2A(,, differs from the homologous calf histone in 61 amino acid positions. aestivum cultivar T 4 has been established from Edman degradation of large overlapping fragments.These differences include an extension of H2A,,, by 19 amino acids at its carboxyl end.Histones in general are considered to play a structural rather than a regulatory role in chromatin [l], however the widespread occurrence of histone variants (isohistones), (for review see [2,31) suggests that variants may modulate the conformation of nucleosomes, and in this way modify their susceptibility to regulatory factors. We have established previously the presence of a number of isohistones of the H2A class in wheat embryos [2, 41. We are reporting now the complete structure of one of these isohistones. This structure constitutes a new histone type in that its C-terminal region is extended beyond the size hitherto considered typical for core histones of that group. MATERIALS AND METHODS Isolation of histone H2A(,,Wheat germ (Triticum aestivum) was obtained from Sasko Mills (Rondebosch, Cape). This material contained at least 80% of the cultivar T 4. Chromatin was isolated essentially according to the method of Johns and Butler [5] with some modifications as described previously [4].Exclusion chromatography of histones was performed at room temperature on a column (4.5 x 80 cm) of Bio-Gel P-60 (100-200 mesh). Approximately 200 mg of total acid-extracted (0.2 M H2S04) histone was dissolved in 10 ml of 6 M urea containing 2% (v/v) 2-mercaptoethanol, loaded onto the column and eluted with 0.02 M HCI (Fig. 1 a).Cation-exchange chromatography was carried out using LKB CM-Trisacryl M equilibrated with 0.05 M sodium acetate adjusted to pH 5.00 with HC1. A linear NaCl gradient was used to elute the H2A(,,-enriched fraction B (Fig. 1 b). Fraction B was then further purified by passage over a BioGel P-60 column equilibrated with 0.02 M HCl, 0.05 M NaCl as described previously [6]. The eluant contained in addition, 10 mM sodium bisulfite, 1 mM PhMeS02F and 1 mM Natosyllysylchloromethane, solubilized first in a small volume of propan-1-01 (Fig. 1 c). The H2A(,,-containing fraction was sometimes recycled over the P-60 column if there was still some contamination evident on gel electrophoresis. Analytical proceduresPolyacrylamide gel electrophoresis (PAGE) employing the detergent Triton X-100 was performed in glass tubes (9 x 0.5 cm) [7, 81.Histone samples were hydrolysed under vacuum for 24 h at 110°C in constant-boiling HCl containing 0.025% (w/v) phenol as antioxidant unless otherwise stated. The analyses were done with either ninhydrin methodology (Beckman 119 automatic analyser) or using hypochlorite and o-phthalaldehyde post-column fluorescence detection (Waters Associates HPLC amino acid analyser [9]).Fragmentation of H2A(,, N-Bromosuccinimide (SucNBr) cleavage [lo] was performed as described previously [ll]. The increase...

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“…Synechococcus PCC 7942 wild type was grown in a basal medium complemented with 4 mM NaNO 3 or 5 mM NH 4 Cl as described previously (1). After membrane preparation, the sample was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) 5 (1).…”

Section: Methodsmentioning

confidence: 99%

The Nitrogen Source‐Dependent 126‐kDa Protein From Synechococcus PCC 7942 Plasmalemma: A Trimer of the NrtA Nitrate‐Binding Protein

Fresneau

Zinovieva²,

Blanot

et al. 2000

IUBMB Life

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Summary Expression of a 126-kDa protein in the plasmalemma (cytoplasmic membrane) fromIn a previous paper (1) we reported the expression of a 126-kDa protein in the plasmalemma from Synechococcus PCC 7942 that is dependent on the nitrogen source as the 45-kDa (NrtA) protein (2-4). Indeed, the 126-kDa protein is absent in ammonium-grown cells, and its quantitative expression is inversely related to the concentration of nitrate or nitrite in the external medium. Moreover, transfer of ammonium-grown cells into a medium containing either nitrate or L-methionine-DLsulfoximine results in expression of this protein. Deletion of the nrtABCD part of the nirA operon, as in the NA3 mutant,

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Column chromatography of amino acids with fluorescence detection

Cited by 295 publications

References 2 publications

Orthotopic liver transplantation for urea cycle enzyme deficiency

Orthotopic liver transplantation for urea cycle enzyme deficiency

The amino acid sequence of wheat histone H2A(1). A core histone with a C-terminal extension

The Nitrogen Source‐Dependent 126‐kDa Protein From Synechococcus PCC 7942 Plasmalemma: A Trimer of the NrtA Nitrate‐Binding Protein

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