High-performance liquid chromatography of Dns-amino acids in the purity control of peptides

Levina, N.B.; Nazimov, Igor V.

doi:10.1016/s0021-9673(01)99187-3

Cited by 18 publications

(4 citation statements)

References 13 publications

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“…At intervals ranging from 10 to 240 min., 50 l of supernatant from perchlorate-treated samples (0.3 M) were removed and subjected to amino acid analysis according to the method of Levina and Nazimov (15). Briefly, the solution was dried in vacuo at 60°C and evaporated to dryness twice from 5 l of water.…”

Section: Methodsmentioning

confidence: 99%

The Asp272–Glu282 Region of Platelet Glycoprotein Ibα Interacts with the Heparin-binding Site of α-Thrombin and Protects the Enzyme from the Heparin-catalyzed Inhibition by Antithrombin III

Cristofaro¹,

Candia²,

Rutella³

et al. 2000

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

The Asp272–Glu282 Region of Platelet Glycoprotein Ibα Interacts with the Heparin-binding Site of α-Thrombin and Protects the Enzyme from the Heparin-catalyzed Inhibition by Antithrombin III

Cristofaro¹,

Candia²,

Rutella³

et al. 2000

Journal of Biological Chemistry

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“…The reaction with dansyl chloride was performed according to Hartley [24], separating and identifying the dansyl derivatives by HPLC [25]. Carboxypeptidase Y (Sigma) was added at a ratio of 1 pg/nmol peptide dissolved in 0.1 M sodium acetate pH 5.5, containing norleucine as an internal standard.…”

Section: Other Methodsmentioning

confidence: 99%

Site‐directed alkylation and site‐site interactions in bovine seminal ribonuclease

Donadio¹,

Tamburrini²,

Donato³

et al. 1986

European Journal of Biochemistry

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Active-site histidine residues of bovine seminal RNase have been found to react with bromoacetic acid and with 2'(3')-O-bromoacetyluridine (BrAcUrd) at a much faster rate than free histidine. The former reagent reacts preferentially at the pros-N of His"', the latter is specific for the tele-N of His". Alkylation with bromoacetic acid is mutually exclusive for either His"' or His" and takes place predominantly at His"', while with BrAcUrd alkylation was found to be selective for His". These results are very similar to those obtained with the same reagents on RNase A, confirming that seminal and pancreatic ribonucleases have similar geometries at their active sites. On the other hand, the kinetics of reaction of bromoacetyluridine with seminal RNase reveal a 'half-of-the sites' reactivity of the enzyme for this reagent, which is found to discriminate between the two structurally identical active sites of the dimeric enzyme.Bovine seminal RNase (BS-RNase) is a dimeric enzyme, which displays non-hyperbolic saturation curves for the substrates of the second, rate-limiting step of the reaction, i.e. the hydrolysis of the pyrimidine nucleoside 2',3'-phosphates (cyclic) produced in the first reaction step [I]. Pancreatic RNase A shares with BS-RNase subunit a perfectly conserved active site, 80% of its primary structure and its three-dimensional structure [2, 31. Recently it has been reported that RNase A also displays non-hyperbolic kinetics when it is artifically dimerized [4].RNase A was the model protein for the pioneering studies of Stein, Moore and their coworkers on active-site mapping, carried out through the use of reagents selectively modifying residues at the active sites of enzymes [5-91. Their results, and those of other research groups [I0 -141, have led to the conclusion that alkylating reagents can selectively react with the active-site histidine residues of RNase A; they can also discriminate between His"', reacting at itspros-N, and His", reacting at its tele-N.The purpose of this study was twofold. We wished to prepare inactive derivatives of BS-RNase through selective modifications of active-site residues. The preparation of such derivatives and their characterization would be of particular Correspondence to G. D'Alessio, Dipartimento di Chimicas, Organica e Biologica, Universita di Napoli, Via Mezzocannone 16, 1-801 34 Napoli, ItalyAbbreviations. BS-RNase, bovine seminal ribonuclease; BrAcUrd, 2'(3')-O-bromoacetyluridine; Cm-His, carboxymethylhistidine; Cm-Cys, carboxymethylcysteine ; HPLC, high-performance liquid chromatography.Enzymes. Ribonuclease, pancreatic (EC 3.1.27.5); ribonuclease, seminal (EC 3.1.27.-).Note. The imidazole N closest to the propionyl side-chain of histidine, formerly designated as N-1, is termed pros, or x , and N-3 is termed tele, or 7 , according to IUPAC-IUB Recommendations [Eur.J. Biochem. 138, 13 (1984)l. value for studying both the peculiar regulatory properties of the enzyme and its interesting biological actions, such as its antispermatogenic [15] and antitumora...

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“…The mixture was allowed to stand for 30 minutes at room temperature in the dark (Levina and Nazimov, 1984;Tapuhi et al 1981) and then loop1 of 2 % ethylamine solution was added. The resulting solution was analyzed by HPLC: column: Nucleosil 5C18, 250 mm x 4.6 mm id.…”

Section: Determination Of L-lysinementioning

confidence: 99%

“…A 100-fold dilution of the above reaction mixture (indirect method) (100 pl) was mixed with 200 pl of lithium carbonate buffer (40 mmol/l, adjusted to pH 9.5 with 1 mol/l HCI) and then loop1 of 1.5 mg/ml dansyl chloride DNS-Cl) acetonitrile solution was added. The mixture was allowed to stand for 30 minutes at room temperature in the dark (Levina and Nazimov, 1984;Tapuhi et al 1981) and then loop1 of 2 % ethylamine solution was added. The resulting solution was analyzed by HPLC: column: Nucleosil 5C18, 250 mm x 4.6 mm id.…”

Section: Determination Of L-lysinementioning

confidence: 99%

Extra‐weak chemiluminescence of drugs. XII. Effect of the molar ratio of amino acid to sugar on extra‐weak chemiluminescence in the Maillard reaction

Kurosaki

Satō

Ishizawa

et al. 1991

J. Biolumin. Chemilumin.

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The extra-weak chemiluminescence in the Maillard reaction caused by the reaction between L-lysine and D-arabinose was measured, and a linear relationship was found between the chemiluminescence and the amount of L-lysine added. After a 1-hour reaction equimolar amounts of D-arabinose and L-lysine were consumed regardless of the initial concentration of D-arabinose. The chemiluminescence of the Maillard reaction originates from Maillard reaction products formed by the equimolar reaction between sugar and amino acid and depends on the concentration of amino acid.

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High-performance liquid chromatography of Dns-amino acids in the purity control of peptides

Cited by 18 publications

References 13 publications

The Asp272–Glu282 Region of Platelet Glycoprotein Ibα Interacts with the Heparin-binding Site of α-Thrombin and Protects the Enzyme from the Heparin-catalyzed Inhibition by Antithrombin III

The Asp272–Glu282 Region of Platelet Glycoprotein Ibα Interacts with the Heparin-binding Site of α-Thrombin and Protects the Enzyme from the Heparin-catalyzed Inhibition by Antithrombin III

Site‐directed alkylation and site‐site interactions in bovine seminal ribonuclease

Extra‐weak chemiluminescence of drugs. XII. Effect of the molar ratio of amino acid to sugar on extra‐weak chemiluminescence in the Maillard reaction

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