Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Crop proteomics: Practical method for high resolution of two-dimensional electrophoresis

작물 단백질체 분석을 위한 이차원 전기영동 사용법

  • Kim, U.G. (Department of Plant Bioscience, Pusan National University) ;
  • Jung, Hwa-Jin (Department of Plant Bioscience, Pusan National University) ;
  • Lee, Su-Ji (Department of Plant Bioscience, Pusan National University) ;
  • Kim, Sun-Tae (Department of Plant Bioscience, Pusan National University)
  • 김유지 (부산대학교 식물생명과학과) ;
  • 정화진 (부산대학교 식물생명과학과) ;
  • 이수지 (부산대학교 식물생명과학과) ;
  • 김선태 (부산대학교 식물생명과학과)
  • Received : 2012.03.10
  • Accepted : 2012.03.20
  • Published : 2012.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Two-dimensional gel electrophoresis (2-DGE) is one of the most important technologies for high-resolution separation of proteins for proteomics. In this study, we present a detail 2-DGE protocol which allows detection and quantification of total plant proteins separated on gels to improve matching in image analysis. This protocol highlighted here may be useful for researchers, who like to first study for the development of protein biomarkers involved in development, biotic and abiotic stresses in plant.

단백질체학에서 이차원 전기영동 (2-DGE)은 고해상도 단백질 분리가 가능한 중요 기술 중 하나이다. 본 논문에서는 2-DGE 이미지 분석 일치도를 향상시킴으로써 작물 단백질 검출과 정량분석이 가능하도록 하는 전반적인 주요 장비, 시약 등을 자세히 기술한 프로토콜을 제시하고 있다. 이 프로토콜은 식물의 발달, 생물학적, 비생물학적 스트레스 반응 등과 관련된 작물 단백질 바이오마커를 개발하기 위한 목적에서 2-DGE를 처음 시도하는 연구자들에게 유용할 것으로 기대한다.

Keywords

References

  1. Agrawal GK, Rakwal R (2011) Rice proteomics: A move toward expanded proteome coverage to comparative and functional proteomics uncovers the mysteries of rice and plant biology. Proteomics 11:1630-1649 https://doi.org/10.1002/pmic.201000696
  2. Carpentier SC, Witters E, Laukens K, Deckers P, Swennen R, Panis B (2005) Preparation of protein extracts from recalcitrant plant tissues: an evaluation of different methods for two‐dimensional gel electrophoresis analysis. Proteomics 5:2497-2507 https://doi.org/10.1002/pmic.200401222
  3. Chen S, Harmon AC (2006) Advances in plant proteomics. Proteomics 6:5504-5516 https://doi.org/10.1002/pmic.200600143
  4. Cho K, Torres NL, Subramanyam S, Deepak SA, Sardesai N, Han O, Williams CE, Ishii H, Iwahashi H, Rakwal R (2006) Protein extraction/solubilization protocol for monocot and dicot plant gel-based proteomics. J Plant Soil 49:413-420
  5. Giavalisco P, Nordhoff E, Lehrach H, Gobom J, Klose J (2003) Extraction of proteins from plant tissues for two‐dimensional electrophoresis analysis. Electrophoresis 24:207-216 https://doi.org/10.1002/elps.200390016
  6. Gorg A, Weiss W, Dunn MJ (2004) Current two-dimensional electrophoresis technology for proteomics. Proteomics 4:3665-3685 https://doi.org/10.1002/pmic.200401031
  7. Gygi SP, Corthals GL, Zhang Y, Rochon Y, Aebersold R (2000) Evaluation of two-dimensional gel electrophoresis- based proteome analysis technology. Proc Natl Acad Sci USA 97:9390-9395 https://doi.org/10.1073/pnas.160270797
  8. Hurkman WJ, Tanaka CK (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol 81(3):802-806 https://doi.org/10.1104/pp.81.3.802
  9. Ideker T, Galistki T, Hood L (2001) A new approach to decoding life: systems biology. Annu Rev Genom Hum G 2:343-372 https://doi.org/10.1146/annurev.genom.2.1.343
  10. Kim ST, Cho KS, Jang YS, Kang KY (2001) Two-dimensional electrophoretic analysis of rice proteins by polyethylene glycol fractionation for protein arrays. Electrophoresis 22: 2103-2109 https://doi.org/10.1002/1522-2683(200106)22:10<2103::AID-ELPS2103>3.0.CO;2-W
  11. Lee Y, Bea S, Seo SG, Shim IS, Kim SH, Kim SG, Kang KY, Kim ST (2011) Korean plant proteomics: pioneers in plant stress physiology. J Plant Biotechnol 38:151-161 https://doi.org/10.5010/JPB.2011.38.2.151
  12. Meyer Y, Grosset J, Chartier Y, Cleyet-Marel JC (1988) Preparation by two-dimensional electrophoresis of proteins for antibody production: Antibodies against proteins whose synthesis is reduced by auxin in tobacco mesophy II protoplasts. Electrophoresis 9:704-712 https://doi.org/10.1002/elps.1150091105
  13. Patterson SD, Aebersold RH (2003) Proteomics: the first decade and beyond. Nat Genet 33:311-323 https://doi.org/10.1038/ng1106
  14. Rabilloud T (2002) Two-dimensional gel electrophoresis in proteomics: Old, old fashioned, but it still climbs up the mountains. Proteomics 2:3-10 https://doi.org/10.1002/1615-9861(200201)2:1<3::AID-PROT3>3.0.CO;2-R
  15. Rabilloud T, Lelong C (2011) Two-dimensional gel electrophoresis in proteomics: A tutorial. J Proteomics 74:1829-1841 https://doi.org/10.1016/j.jprot.2011.05.040
  16. Saravanan RS, Rose JKC (2004) A critical evaluation of sample extraction techniques for enhanced proteomic analysis of recalcitrant plant tissues. Proteomics 4:2522-2532 https://doi.org/10.1002/pmic.200300789
  17. Wang W, Scali M, Vignani R, Spadafora A, Sensi E, Mazzuca S, Cresti M (2003) Protein extraction for two‐dimensional electrophoresis from olive leaf, a plant tissue containing high levels of interfering compounds. Electrophoresis 24:2369-2375 https://doi.org/10.1002/elps.200305500
  18. Wittmann‐Liebold B, Graack HR, Pohl T (2006) Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry. Proteomics 6:4688-4703 https://doi.org/10.1002/pmic.200500874

Cited by

  1. Abundant storage protein depletion from tuber proteins using ethanol precipitation method: Suitability to proteomics study vol.15, pp.10, 2015, https://doi.org/10.1002/pmic.201400526
  2. Depletion of abundant plant RuBisCO protein using the protamine sulfate precipitation method vol.13, pp.14, 2013, https://doi.org/10.1002/pmic.201200555
  3. Development of Proteomics-based Biomarkers for 4 Korean Cultivars of Sorghum Seeds (Sorghum bicolor (L.) Moench) vol.32, pp.1, 2013, https://doi.org/10.5338/KJEA.2013.32.1.48
  4. An Integrated Biochemical, Proteomics, and Metabolomics Approach for Supporting Medicinal Value of Panax ginseng Fruits vol.7, 2016, https://doi.org/10.3389/fpls.2016.00994
  5. Proteomics Analysis of Early Salt-Responsive Proteins in Ginseng (Panax ginseng C. A. Meyer) Leaves vol.22, pp.5, 2014, https://doi.org/10.7783/KJMCS.2014.22.5.398