Quantitative ultramicroanalysis of amino acids in the form of their DNS-derivatives

Spivak, V.A.; Shcherbukhin, Vladimir V.; Орлов, В. М.; Varshavsky, Ja.M.

doi:10.1016/0003-2697(71)90415-5

Cited by 32 publications

(7 citation statements)

References 8 publications

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“…Dansylation is a well‐known derivatization technique used in the past for quantitative determination of amino acids 12–17. Dansyl chloride reacts primarily with primary amines at alkaline pH (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We were interested in using UV‐absorbing dansyl chloride, a reagent used in the past for derivatization of amino acids, to derivatize peptides to make them more readily detectable. Dansyl chloride is known to react with the ε ‐amino group of lysine and the hydroxyl group of tyrosine as well as the N‐terminal amino group 12–17. Thus it might be possible to observe lysine‐ and tyrosine‐containing tryptic peptides with different numbers of dansyl groups, which could help identify the protein with greater confidence.…”

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confidence: 99%

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Dansylation of tryptic peptides for increased sequence coverage in protein identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric peptide mass fingerprinting

Park

Song

Kim

2005

Rapid Comm Mass Spectrometry

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A database search using peptide mass fingerprints obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry leads to protein identification with incomplete sequence coverage, because certain peptides are preferentially desorbed/ionized and some are not detected at all. We show that certain tryptic peptides mainly with C-terminal arginine not detected before derivatization become detectable upon dansylation. Others, mainly with C-terminal lysine, are suppressed. An increase in protein sequence coverage and protein identification score by combined data from underivatized and dansylated peptides in database search is demonstrated using human amnion proteins (human serum albumin precursor, calmodulin, collagen alpha 2(VI) chain precursor, galectin-3) separated by two-dimensional gel electrophoresis as well as femtomole amounts of BSA in solution.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Dansylation of tryptic peptides for increased sequence coverage in protein identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric peptide mass fingerprinting

Park

Song

Kim

2005

Rapid Comm Mass Spectrometry

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“…Also the pyridoxyl derivatives of amino acids (3) were not considered since they, too, are separated on ion exchangers. The l-A/./V'-dimethylaminonaphthalene-S-sulfonyl derivatives of amino groups (dansyl derivatives) show very strong fluorescence and already have been used in protein chemistry (4,5) and in thin-layer chromatography (6)(7)(8). The preparation of these derivatives is fast and quantitative (9), and their strong fluoresdence enables detection at concentrations at least four to five orders of magnitude lower than is possible with the ninhydrin reaction.…”

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confidence: 99%

Separation of amino acids by high performance liquid chromatography

Bayer¹,

Grom²,

Kalteneger³

et al. 1976

Anal. Chem.

196

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“…Dansyl amino acids were separated by thin-layer chromatography (Spivak et al, 1971) and their concentration determined by spectrofluorimetry, according to Gros and Labouesse (1969). Amino acid analysis after 24 h hydrolysis in 3 N p-toluene-sulfonic acid, was performed in a Kontron amino acid analyzer (Durrum DC-4 resin), equipped for the fluorimetric detection of amino acids.…”

Section: End Group and Amino Acid Analysismentioning

confidence: 99%

Transient conformational states in proteins followed by differential labeling

Ghélis¹

1980

Biophysical Journal

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Refolding of previously denatured and reduced elastase has been followed by titration of chemical reactivities of amino acid side chains to study the topography of the protein in the native state, and the microenvironment variations of protein side chains during the structural transition. Groups accessible to chemical reagents in the denatured form and buried in the "native" form were used as a local conformational probe. Times of labeling, depending on the reagent used, ranged from 100 to 800 ms. The reaction was stopped by isotopic dilution with an excess of unlabeled reagent under denaturing conditions to obtain a chemically homogeneous but heterogeneously labeled material. Peptide fractionation after degradation of the labeled proteins allowed the determination of the amount of radioactive label incorporated by the individual side chains during the refolding. Refolding rates, determined by physicochemical, enzymatic or immunochemical criteria, were compared with the conformational states of protein areas and evaluated by the variation of chemical reactivity at various denaturant concentrations. The importance of the last folding stages is emphasized by the results obtained which indicate that early during the refolding, two domain substructures (H-40 to H-71 and M-180 to H-200)( are stabilized, while the protein remains inactive at the time ranges of the labeling reactions.

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Quantitative ultramicroanalysis of amino acids in the form of their DNS-derivatives

Cited by 32 publications

References 8 publications

Dansylation of tryptic peptides for increased sequence coverage in protein identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric peptide mass fingerprinting

Dansylation of tryptic peptides for increased sequence coverage in protein identification by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometric peptide mass fingerprinting

Separation of amino acids by high performance liquid chromatography

Transient conformational states in proteins followed by differential labeling

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