Noninterfering Synthetic Peptides as Internal Controls for Amino Acid Sequencing of Sample Unknowns

Noninterfering synthetic peptides have been designed that may be used as internal sequencing standards (ISS-1 and ISS-2) by placing them in an amino acid sequencer with the sample. The peptides are composed of four unnatural amino acids which all yield phenylthiohydantoin derivatives having unique r...

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Bibliographic Details
Published in:Analytical biochemistry 1994-05, Vol.219 (1), p.71-81
Main Authors: Parmelee, D.C., Hoang, T.N., Benjamin, T., Sechi, S.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino Acids - analysis
Analytical biochemistry: general aspects, technics, instrumentation
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Biotechnology
Chromatography, High Pressure Liquid - methods
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Indicators and Reagents
Methods. Procedures. Technologies
Peptides - chemical synthesis
Peptides - chemistry
Peptides - isolation & purification
Reference Standards
Spectrometry, Mass, Fast Atom Bombardment - methods
Synthetic digonucleotides and genes. Sequencing
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Summary:Noninterfering synthetic peptides have been designed that may be used as internal sequencing standards (ISS-1 and ISS-2) by placing them in an amino acid sequencer with the sample. The peptides are composed of four unnatural amino acids which all yield phenylthiohydantoin derivatives having unique retention times compared with those obtained from the commonly observed natural residues. These internal standards indicate how the entire sequencing system was functioning during the actual analysis of an unknown by providing an initial yield and numerous repetitive yields. Verifying proper operation is extremely important when cycles appear blank due to the presence of modified amino acids or a blocked N-terminus. In addition, the ISS peptides can detect and identify different types of sequencing errors. This sometimes eliminates the need to rerun a sample due to blank cycles caused by mechanical malfunctions which result in failure to cleave the N-terminal residues. ISS-1 or ISS-2 may also be utilized during method development to compare different sample supports and to normalize sequencing data from proteins or peptides that have been treated differently.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1994.1233