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The enhancement of protein separation by duplex SDS-PAGE

Duplex SDS-PAGE를 이용한 단백질 분리향상

  • Pyo, Jae Sung (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • Roh, Si Hun (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • Song, Jin-Su (Department of Chemistry, Seoul National University) ;
  • Lee, Kyung Hyeon (Department of Chemistry, Seoul National University) ;
  • Kim, Hie-Joon (Department of Chemistry, Seoul National University) ;
  • Park, Jeong Hill (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • Kwon, Sung Won (College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University)
  • Received : 2006.09.22
  • Accepted : 2006.11.20
  • Published : 2006.12.28

Abstract

The protein separation with molecular weight using SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is the one of the most conventional and simple techniques. In, this study, two dimensional SDS-PAGE using same separation principle consecutively was investigated and compared with one dimensional SDS-PAGE. The enhanced separation from duplex SDS-PAGE was observed and separated proteins in the gel were identified by MALDI TOF MS. Identified proteins from different gel spots were found to have different gi numbers. Therefore, duplex SDS-PAGE separation method will be used for economic separation method in the future because only tiny amount of inexpensive reagents are used to perform duplex SDS-PAGE.

SDS-PAGE를 이용한 일반적인 단백질 분리는 단백질을 분자량에 따라 분리하는 방법이며 가장 보편적이고, 간단한 방법 중의 하나이다. 본 연구는 SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)를 두 번에 걸쳐 동일한 분리 원리로 이차원으로 전개하는 방법의 전기영동을 시도하여 기존의 일차원 전기영동 분석법과 비교하였으며, 그 결과 향상된 분리능이 본 연구의 이차원 전기영동법에서 확인되었다. Gel에서 분리된 단백질들은 MALDI TOF MS를 이용하여 동정하여 서로 다른 단백질임이 확인되었으며, 이러한 duplex SDS-PAGE 분리법은 상대적으로 적은 비용으로 단백질 분리능을 용이하게 향상시킬 수 있는 경제적인 분석법으로 이용될 수 있을 것이다.

Keywords

References

  1. P. Jungblut and B. Wittmann-Liebold, J. Biotechnology, 41, 111-120(1995) https://doi.org/10.1016/0168-1656(95)00006-C
  2. K. Kahn, Science, 270, 369-370(1995) https://doi.org/10.1126/science.270.5234.270
  3. M. Norin and M. Sundstrom, Trends Biotechnol., 20, 79-84(2002) https://doi.org/10.1016/S0167-7799(01)01884-4
  4. A. Pandey and M. Mann, Nature, 405, 837-846(2000) https://doi.org/10.1038/35013174
  5. K. Einer-Jensen, T. N. Krogh, P. Roepstorff, N. Lorenzen, J. Virology, 72, 10189-10196(1998)
  6. D. Ungar, T. Oka, E. E. Brittle, E. Vasile, V. V. Lupashin, J. E. Chatterton, J. E. Heuser, M. Krieger, M. G. Waters, J. Cell Biol., 157, 405-415(2002) https://doi.org/10.1083/jcb.200202016
  7. U. K. Laemmli, Nature, 227, 680-685(1970) https://doi.org/10.1038/227680a0
  8. J. Kim, J. Kim, H. Kim, Anal. Chem., 77(22), 7483- 7488(2005) https://doi.org/10.1021/ac051152l