Micropreparative separation of peptides derived from sodium dodecyl sulphate-solubilized proteins
| Autor: | Armin Bosserhoff, Jochen Wallach, Rainer Frank |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
Sodium
Molecular Sequence Data chemistry.chemical_element Alcohol Peptide Biochemistry High-performance liquid chromatography Analytical Chemistry chemistry.chemical_compound Amino Acid Sequence Bovine serum albumin Fragmentation (cell biology) Integral membrane protein Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography biology Organic Chemistry Sodium Dodecyl Sulfate Bacteriorhodopsin General Medicine chemistry Bacteriorhodopsins biology.protein Indicators and Reagents Spectrophotometry Ultraviolet Peptides |
| Zdroj: | Journal of Chromatography A. 473:71-77 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/s0021-9673(00)91291-3 |
| Popis: | A systematic investigation of the influence of the detergent sodium dodecyl sulphate (SDS) on micropreparative peptide separations on microbore reversed-phase high-performance liquid chromatographic columns is reported. A tryptic digest of bovine serum albumin and a mixture of synthetic peptides were used to monitor the separation behaviour of a 1.6 mm I.D. Nucleosil C18 column in the presence of various amounts of SDS. The data demonstrate that even traces of SDS in the sample reduce the separation efficiency and peptide recovery. An extraction method is presented which reduces the SDS content in peptide mixtures below the critical concentration without affecting significantly the recovery of individual peptides. After acidification of the sample, the detergent is extracted into heptane-isoamyl alcohol (4:1, v/v). In combination with chemical or enzymatic fragmentation techniques, this extraction method facilitates the sequence analysis of minute amounts of SDS-solubilized hydrophobic proteins. The applicability of the method is demonstrated on the example of the integral membrane protein bacteriorhodopsin. |
| Databáze: | OpenAIRE |
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