Abstract
A manual high-sensitivity sequencing method is described, in which 4-NN-dimethylaminoazobenzene 4'-isothiocyanate is used for the stepwise degradation of amino acid residues from the peptides. The 4-NN-dimethylaminoazobenzene 4'-thiazolinones of amino acids that were released, after conversion into their thiohydantoin derivatives, were identified by t.l.c. on polyamide sheets. This new method is simple and sensitive, and requires only 2-10nmol of peptides or proteins for extended sequence analysis. The method was tested on the sequence analysis of a hexapeptide (Leu-Trp-Met-Arg-Phe-Ala), bradykinin, glucagon and native lysozyme. Results show that the proposed procedure is a sensitive method for the sequence determination of short peptides as well as for the partial sequence determination of intact proteins.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barrett G. C., Leigh P. H. Rapid aminolysis of benzoylthionocholines, applied in peptide sequence analysis. Relevance to physiological properties of sulphur analogues of choline esters. FEBS Lett. 1975 Sep 1;57(1):19–21. doi: 10.1016/0014-5793(75)80142-6. [DOI] [PubMed] [Google Scholar]
- Bradbury J. H., Crompton M. W., Warren B. Determination of the sequence of peptides by PMR spectroscopy. Anal Biochem. 1974 Nov;62(1):310–316. doi: 10.1016/0003-2697(74)90394-7. [DOI] [PubMed] [Google Scholar]
- Bridgen J. High sensitivity amino acid sequence determination. Application to proteins eluted from polyacrylamide gels. Biochemistry. 1976 Aug 10;15(16):3600–3604. doi: 10.1021/bi00661a030. [DOI] [PubMed] [Google Scholar]
- Chang J. Y., Creaser E. H. A novel manual method for protein-sequence analysis. Biochem J. 1976 Jul 1;157(1):77–85. doi: 10.1042/bj1570077. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang J. Y., Creaser E. H., Bentley K. W. 4-NN-dimethylaminoazobenzene 4'-isothiocyanate, a new chromophoric reagent for protein sequence analysis. Biochem J. 1976 Mar 1;153(3):607–611. doi: 10.1042/bj1530607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang J. Y., Creaser E. H. Improved chromatographic identification of coloured amino acid thiohydantoins. J Chromatogr. 1977 Feb 11;132(2):303–307. doi: 10.1016/s0021-9673(00)89304-8. [DOI] [PubMed] [Google Scholar]
- Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
- Laursen R. A. Solid-phase Edman degradation. An automatic peptide sequencer. Eur J Biochem. 1971 May 11;20(1):89–102. doi: 10.1111/j.1432-1033.1971.tb01366.x. [DOI] [PubMed] [Google Scholar]
- Morris H. R., Williams D. H., Ambler R. P. Determination of the sequences of protein-derived peptides and peptide mixtures by mass spectrometry. Biochem J. 1971 Nov;125(1):189–201. doi: 10.1042/bj1250189. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Previero A., Gourdol A., Derancourt J., Coletti-Previero M. A. Base catalyzed aminolysis of carbodithioic esters and its interest in solid phase sequential analysis of peptides. FEBS Lett. 1975 Mar 1;51(1):68–72. doi: 10.1016/0014-5793(75)80856-8. [DOI] [PubMed] [Google Scholar]
- Priddle J. D., Rose K., Offord R. E. The separation and sequencing of permethylated peptides by mass spectrometry directly coupled to gas-liquid chromatography. Biochem J. 1976 Sep 1;157(3):777–780. doi: 10.1042/bj1570777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tarr G. E. A general procedure for the manual sequencing of small quantities of peptides. Anal Biochem. 1975 Feb;63(2):361–370. doi: 10.1016/0003-2697(75)90358-9. [DOI] [PubMed] [Google Scholar]
- Weiner A. M., Platt T., Weber K. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J Biol Chem. 1972 May 25;247(10):3242–3251. [PubMed] [Google Scholar]