Abstract

Peptide sequencing remains one of the most important analytical problems in molecular biology. Automated Edman degradation is established as a routine technique in most protein bio-chemical core facilities, and is a straightforward method to obtain enough sequence information to clone a gene [1, 2]. At sufficiently high sample amounts data interpretation is easy and can even be automated. At small sample amounts of less than about 2 picomoles of starting peptide material, however, sequencing by Edman degradation cam become ambiguous. The first and last cycles in an analysis may not be assignable because of background amino acids and because of sample wash out, respectively. Several amino acids such as His, Cys, Trp and Arg are hardly detectable because of low recoveries and low detection yields. An inherent feature of Edman degradation is the need for a pure peptide fraction, as single amino acids are detected in every cycle. This peptide purity is increasingly difficult to attain as sample levels become lower and re-chromatography is not an option.

Keywords

Edman degradationPeptideAmino acidProtein sequencingSample (material)Degradation (telecommunications)ChromatographyPeptide sequenceAmino acid analysisChemistrySequence (biology)Computational biologyBiochemistryBiologyComputer scienceGene

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Publication Info

Year
1996
Type
book-chapter
Pages
245-265
Citations
13
Access
Closed

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Cite This

Matthias Wilm, Tony Houthaeve, Gert Talbo et al. (1996). Approaches to the Practical Use of MS/MS in a Protein Sequencing Facility. Humana Press eBooks , 245-265. https://doi.org/10.1007/978-1-4612-0229-5_13

Identifiers

DOI
10.1007/978-1-4612-0229-5_13
PMID
41262706
PMCID
PMC12623507

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Data completeness: 77%