05SAR-PAGE: Separation of protein dimerization and modification using a gel with 0.05% sarkosyl

Anal Chim Acta. 2020 Mar 8:1101:193-198. doi: 10.1016/j.aca.2019.12.013. Epub 2019 Dec 9.

Abstract

A protein gel electrophoresis procedure using 0.05% w/v sarkosyl, is reported. The method called 05SAR-PAGE can be used to identify the native masses, dimeric states and modification states of proteins, and also be suitable for pursuing native electroblotting and immunodetection. It has been demonstrated by NMR spectroscopy that 0.05% w/v SAR is much milder than SDS, so it has subtle effects on the native structure of proteins. Therefore, the non-covalent dimerization of PhoBN and PhoRcp can be identified by 05SAR-PAGE which cannot be observed by SDS-PAGE. It has also been demonstrated that 05SAR-PAGE can be used to identify the phosphorylated or methylated proteins. Besides, 05SAR-PAGE shows the advantages of simple operation and low cost, and can be easily adapted to diverse applications.

Keywords: Dimerization; Gel electrophoresis; Immunodetection; Modification states; NMR spectroscopy; Sarkosyl.

MeSH terms

  • Bacterial Proteins / analysis*
  • Bacterial Proteins / chemistry
  • Electrophoresis, Polyacrylamide Gel / methods
  • Escherichia coli / chemistry
  • Escherichia coli Proteins / analysis*
  • Escherichia coli Proteins / chemistry
  • Protein Multimerization
  • Sarcosine / analogs & derivatives*
  • Sarcosine / chemistry
  • Transcription Factors / analysis*
  • Transcription Factors / chemistry

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • PhoB protein, E coli
  • Transcription Factors
  • PhoR protein, Bacteria
  • sarkosyl
  • Sarcosine