PMID:12071708
| Citation |
Arcidiacono, S, Butler, MM and Mello, CM (2002) A rapid selective extraction procedure for the outer membrane protein (OmpF) from Escherichia coli. Protein Expr. Purif. 25:134-7 |
|---|---|
| Abstract |
Porins are essential pore-forming proteins found in the outer membrane of several gram-negative bacteria. Investigating the relationships between molecular structure and function involves an extremely time-consuming and labor-intensive purification procedure. We report a method for rapid extraction of the outer membrane protein, OmpF, from freeze-dried Escherichia coli cells using valeric acid, alleviating the effort and time in sample preparation. Extraction results in a highly enriched fraction containing OmpF as 76% of the total protein content. The apparent molecular mass determined by SDS-PAGE mobility was 38,900, similar to that of the monomeric form of OmpF. N-terminal sequencing yielded 23 amino acids with 100% identity to the published OmpF sequence. The trimeric form of OmpF was observed in unheated samples run on SDS-PAGE and analysis of these samples by periodic acid/silver staining revealed the presence of unbound lipopolysaccharides. Furthermore, this method should prove useful for isolating other outer membrane proteins. |
| Links |
PubMed Online version:10.1006/prep.2002.1619 |
| Keywords |
Amino Acid Sequence; Chemistry Techniques, Analytical/methods; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Escherichia coli/metabolism; Freezing; Hydrogen-Ion Concentration; Lipopolysaccharides/pharmacology; Molecular Sequence Data; Pentanoic Acids/pharmacology; Porins/chemistry; Porins/isolation & purification; Protein Structure, Tertiary; Sequence Homology, Amino Acid |
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