Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure

We have examined two‐dimensional electrophoresis (2‐DE) gel maps of polypeptides from the Gram‐negative bacterium Methylococcus capsulatus (Bath) and found the same widespread trains of spots as often reported in 2‐DE gels of polypeptides of other Gram‐negative bacteria. Some of the trains of polype...

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Veröffentlicht in:Electrophoresis 2003-02, Vol.24 (4), p.757-761
Hauptverfasser: Berven, Frode S., Karlsen, Odd A., Murrell, J. Colin, Jensen, Harald B.
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container_issue 4
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container_title Electrophoresis
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creator Berven, Frode S.
Karlsen, Odd A.
Murrell, J. Colin
Jensen, Harald B.
description We have examined two‐dimensional electrophoresis (2‐DE) gel maps of polypeptides from the Gram‐negative bacterium Methylococcus capsulatus (Bath) and found the same widespread trains of spots as often reported in 2‐DE gels of polypeptides of other Gram‐negative bacteria. Some of the trains of polypeptides, both from the outer membrane and soluble protein fraction, were shown to be generated during the separation procedure of 2‐DE, and not by covalent post‐translational modifications. The trains were found to be regenerated when rerunning individual polypeptide spots. The polypeptides analysed giving this type of trains were all found to be classified as stable polypeptides according to the instability index of Guruprasad et al. (Protein Eng. 1990, 4, 155–161). The phenomenon most likely reflects conformational equilibria of polypeptides arising from the experimental conditions used, and is a clear drawback of the standard 2‐DE procedure, making the gel picture unnecessarily complex to analyse.
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Some of the trains of polypeptides, both from the outer membrane and soluble protein fraction, were shown to be generated during the separation procedure of 2‐DE, and not by covalent post‐translational modifications. The trains were found to be regenerated when rerunning individual polypeptide spots. The polypeptides analysed giving this type of trains were all found to be classified as stable polypeptides according to the instability index of Guruprasad et al. (Protein Eng. 1990, 4, 155–161). The phenomenon most likely reflects conformational equilibria of polypeptides arising from the experimental conditions used, and is a clear drawback of the standard 2‐DE procedure, making the gel picture unnecessarily complex to analyse.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>12601748</pmid><doi>10.1002/elps.200390091</doi><tpages>5</tpages></addata></record>
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subjects Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Conformational equilibria
Electrophoresis, Gel, Two-Dimensional - methods
Methylococcus capsulatus - metabolism
Post-translational modifications
Proteomics
Trains of spots
Two-dimensional gel electrophoresis
title Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure
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