KOREAN JOURNAL OF CROP SCIENCE (한국작물학회지)

The Korean Society of Crop Science (한국작물학회)
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A New Removal Method of Glutelin Storage Proteins for the Proteome Study of Non-Glutelin Proteins in Rice Seeds

벼종자 미랑 단백질의 프로테오믹스 연구를 위한 글루테린 저장 단백질의 제거방법

  • Woo, Sun-Hee (Department of Crop Science, Chungbuk National University) ;
  • Kim, Se-Young (Proteomics Team, Korea Basic Science Institute) ;
  • Kim, Tae-Seon (Department of Crop Science, Chungbuk National University) ;
  • Cho, Seong-Woo (Department of Crop Science, Chungbuk National University) ;
  • Cho, Kun (Proteomics Team, Korea Basic Science Institute) ;
  • Chung, Keun-Yook (Department of Agricultural Chemistry, Chungbuk National University) ;
  • Kim, Sun-Lim (National Institute of Crop Science) ;
  • Cho, Yong-Gu (Department of Crop Science, Chungbuk National University) ;
  • Kim, Hong-Sig (Department of Crop Science, Chungbuk National University) ;
  • Song, Boem-Heon (Department of Crop Science, Chungbuk National University) ;
  • Lee, Chul-Won (Department of Crop Science, Chungbuk National University) ;
  • Jong, Seung-Keun (Department of Crop Science, Chungbuk National University) ;
  • Park, Young-Mok (Proteomics Team, Korea Basic Science Institute)
  • Published : 2006.11.30
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Abstract

Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.

본 연구는 벼종자 미량 단백질의 프로테오믹스 연구를 위하여 벼종자에 고 함량으로 존재하는 벼종자 글루테린 저장 단백질을 제거하는 방법에 관한 것이다. 따라서 본 연구는, (A) 벼종자에 액체 질소를 가하고 분쇄하여 벼종자 가루를 만드는 분쇄단계; (B) 상기 분쇄된 벼종자 가루를 물에 현탁하여 현탁액을 만드는 현탁단계; (C)상기 현탁액 중 미용해 물질을 제거하는 분리단계를 포함하는, 벼종자 미량 단백질의 프로테오믹스 연구를 위한 벼종자 글루테린 저장 단백질의 제거방법에 관하여 검토하였다. 본 연구의 결과, 단순하고 신속하며 저렴하고 효율적인 방법으로 미량 비글루테린 단백질들을 용이하게 동정할 수 있을 것으로 판단되었다.

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