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Proteins Edman technique

Fatty acylation of proteins is a common posttranslational modification that blocks sequencing of the N-terminal amino acids by Edman techniques. Mass spectrometry can play an essential role in such sequencing, but it is not always trivial to locate the blocked N-terminal peptide. Besides acetylation, acylation of amino acids with longer-chain fatty acids (e.g., myristic acid) has been reported. [Pg.113]

The Edman degradation, a process by which amino acids are removed stepwise from the NH 2-terminus of peptides and proteins, has evolved since its introduction (Edman, 1950) to the point where it is currently the most valuable tool for protein sequence analysis. Several innovations have led to its widespread use. One of the first was the dansyl Edman technique (Gray, 1967), which is useful for sequencing at the nanomole level. Accelerated manual procedures have been developed by Niall and Potts (1970) and Tarr (1975). During the past decade the rate of sequencing has been... [Pg.261]

The ability to visualize spots on a 2D gel, while useful as a fingerprint, is not the same as protein identification. Protein sequencing by the Edman degradation technique is the classical means of determining... [Pg.11]

N-terminal sequencing is normally undertaken by Edman degradation (Figure 7.5). Although this technique was developed in the 1950s, advances in analytical methodologies now facilitate fast and automated determination of up to the first 100 amino acids from the N-terminus of most proteins, and usually requires a sample size of less than 1 umol to do so (Figure 7.6). [Pg.188]

Ingenious protein sequenators have been devised to carry out the Edman degradation automatically.242 244-246 Each released phenylthiohydantoin is then identified by HPLC or other techniques. Commercial sequenators have often required 5-20 nmol of peptide but new microsequenators can be used with amounts as low as 5-10 picomoles or less.247 248... [Pg.118]

The formation of methyl- (MTH) or phenylthiohydantoin (PTH) amino acids is a valuable technique for sequencing of amino acids in peptides and proteins by the Edman degradation procedure [1], HPLC is very useful for the separation of MTH- or PTH-amino acids as adsorption [2,3], reversed-phase [4] and ion-exchange [S] chromatography. [Pg.113]

This technique gives information about the protein s primary structure, which may include its amino and/or carboxyl terminal groups (Edman, 1950). For recombinant DNA-derived proteins, this analysis serves to confirm the amino acid sequence predicted by the DNA sequence. The analysis can also be useful to determine the protein s homogeneity. [Pg.337]

Increase in sensitivity and efficiency of analysis in structural studies of enzymes with a gas phase sequencer have made it possible to determine the primary structure in a shorter period of time with a small amount of enzyme at the picomole or even femtomole level. In addition, thanks to the DNA sequencing technique the number of enzymes (or proteins) whose amino acid sequences are registered in a data base file has expanded explosively. The introduction of mass spectrometry on the primary structure determination of protein has stimulated the search for a new methodology other than Edman chemistry. [Pg.14]

Today s sequence techniques have achieved 9-10 orders of magnitude higher detection limit for PTH-amino acid than Edman s first report, in which he described the sequence of Ala-Leu-Gly with 100 pmol. Matsuo et al. succeeded in determing the structure of FSH-RH (LH-RH), a decapeptide hormone, in 1971 by using the Dansyl Edman method.83 This historical work used 200 nmol of FSH-RH purified from 165,000 porcine hypothalami and led a principal investigator, A. Schally, to win the Nobel Prize. Today, we would only need less than one hundred porcine hypothalami to complete the same job on a state-of-art sequencer. Those who plan the structural analysis of even barely detectable amounts of a peptide or protein are urged to start the plan such study could open up a new area in biochemistry. [Pg.35]

The primary structure (i.e., the amino acid sequence) of a protein can be determined by stepwise chemical degradation of the purified protein. By far the most powerful and commonly used technique for doing this is the automated Edman degradation. The amino terminal amino acid residue of the polypeptide is reacted with Edman s reagent (phenylisothiocyanate) to form the phenylthiocar-bamyl derivative, which is removed without hydrolysis of the other peptide bonds by cyclization in anhydrous acid. The amino acid derivative is converted to the more stable phenylthiohydantoin and identified by HPLC. The process can be repeated many times, removing the amino acids from the amino terminus of the polypeptide one residue at a time and identifying them until the entire sequence... [Pg.86]

MALDI-TOF-MS and Edman sequencing have become complementary techniques in protein structural characterization. Ideally, direct collection of peptide fractions should be to a support that... [Pg.169]


See other pages where Proteins Edman technique is mentioned: [Pg.7]    [Pg.184]    [Pg.122]    [Pg.527]    [Pg.331]    [Pg.333]    [Pg.1029]    [Pg.186]    [Pg.35]    [Pg.190]    [Pg.23]    [Pg.205]    [Pg.5]    [Pg.690]    [Pg.34]    [Pg.67]    [Pg.231]    [Pg.457]    [Pg.231]    [Pg.232]    [Pg.457]    [Pg.654]    [Pg.14]    [Pg.70]    [Pg.576]    [Pg.65]    [Pg.17]    [Pg.285]    [Pg.17]    [Pg.29]    [Pg.314]    [Pg.1029]    [Pg.123]    [Pg.267]    [Pg.89]    [Pg.169]    [Pg.38]    [Pg.4]    [Pg.4]   
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Edman technique

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