KOREAN JOURNAL OF CROP SCIENCE (한국작물학회지)

The Korean Society of Crop Science (한국작물학회)
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Optimization of SELDI-TOF MS for Peptide Profiling of Sorghum Seed

수수종자의 펩타이드 분석을 위한 SELDI-TOF MS 최적화 연구

  • Park, Sei Joon (Institute of Ecological Phytochemistry, Hankyong National University) ;
  • Park, June Young (Department of Plant Life and Environmental Science, Hankyong National University) ;
  • Lee, Yong Ho (Institute of Ecological Phytochemistry, Hankyong National University) ;
  • Hwang, Su Min (Department of Plant Life and Environmental Science, Hankyong National University) ;
  • Kim, A Ram (Department of Plant Life and Environmental Science, Hankyong National University) ;
  • Ko, Jee-Yeon (Department of Functional Crop, NICS, RDA) ;
  • Kim, Tae Wan (Institute of Ecological Phytochemistry, Hankyong National University)
  • 박세준 (국립한경대학교 식물생태화학연구소) ;
  • 박준영 (국립한경대학교 식물생명환경과학과) ;
  • 이용호 (국립한경대학교 식물생태화학연구소) ;
  • 황수민 (국립한경대학교 식물생명환경과학과) ;
  • 김아람 (국립한경대학교 식물생명환경과학과) ;
  • 고지연 (농촌진흥청 국립식량과학원 기능성작물부) ;
  • 김태완 (국립한경대학교 식물생태화학연구소)
  • Received : 2012.10.15
  • Accepted : 2013.01.21
  • Published : 2013.03.31
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Abstract

For accurate analysis of low molecular peptides using SELDI-TOF MS (surface enhanced laser desorption/ionization time of flight mass spectrometry), the optimized analytical conditions should be established for a specific biological sample. This study was conducted to optimize SELDI-TOF MS analytical conditions for profiling low molecular peptide below 10 kDa presented in sorghum seeds. Analytical conditions were as follows; (1) protein chips: CM10 (weak cation exchanger) and Q10 (strong anion exchanger), (2) dilution factors of binding buffer: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, and 1/200, (3) the stringency of Q10 binding buffer: 10 mM and 100 mM, and (4) protein extraction buffers: sodium borate, sodium borate + acetone, phenol, and TCA buffers. Optimum dilution factors were selected as 1/20 and 1/50 in both protein chips, CM10 and Q10. Low stringency of Q10 binding buffer (10mM) detected more peptide peaks than high stringency (100 mM). Selected protein extraction buffers of sorghum seed for SELDI-TOF MS analysis was the sodium borate buffer in the range of 2~10 kDa, while the phenol buffer was more suitable in the range of 10~20 kDa.

SELDI-TOF MS를 활용한 저분자 펩타이드 분석을 위해서는 분석시료에 대한 최적화 분석조건을 확립하는 것이 필수적으로 선행되어야 한다. 본 연구는 수수 종자 내 존재하는 10 kDa 이하의 저분자 펩타이드를 프로파일링하기 위하여 활용된 SELDI-TOF MS의 최적화 분석조건을 확립하는데 있다. 분석조건은 (1) 프로테인 칩: CM10(weak cation exchanger), Q10(strong anion exchanger), (2) 바인딩 버퍼의 희석배수: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, 1/200, (3) Q10의 바인딩 버퍼 강도: 10 mM, 100 mM, (4) 단백질 추출버퍼: sodium borate, sodium borate + acetone, phenol, TCA 버퍼로 하였다. 1. 바인딩 버퍼의 희석배수는 CM10과 Q10 모두 1/20과 1/50이 최적화로 나타났다. 2. Q10의 바인딩 버퍼 강도는 농도가 약한 10 mM에서 더 많은 피크가 검출되었다. 3. SELDI-TOF MS 분석에 적합한 수수 종자단백질 추출 버퍼로는 2~10 kDa 범위에서는 sodium borate 버퍼와 10~20 kDa 범위에서는 phenol 버퍼로 분석되었다.

Keywords

References

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