DOI QR코드

DOI QR Code

Analysis of Protein Function and Comparison on Expression of Protein in Taekwang During Maturation using Proteomic Techniques

Proteomics를 이용한 등숙기 차이에 따른 콩 종실 저장단백질 발현양상 비교 분석

  • Cho, Seong-Woo (Crop Breeding Research Division, NICS, RDA) ;
  • Kim, Tae-Sun (Department of Crop Science, Chungbuk National University) ;
  • Kwon, Soo-Jeong (Department of Crop Science, Chungbuk National University) ;
  • Roy, Swapan Kumar (Department of Crop Science, Chungbuk National University) ;
  • Lee, Chul-Won (Department of Crop Science, Chungbuk National University) ;
  • Kim, Hong-Sig (Department of Crop Science, Chungbuk National University) ;
  • Woo, Sun-Hee (Department of Crop Science, Chungbuk National University)
  • 조성우 (국립식량과학원 작물육종과) ;
  • 김태선 (충북대학교 식물자원학과) ;
  • 권수정 (충북대학교 식물자원학과) ;
  • ;
  • 이철원 (충북대학교 식물자원학과) ;
  • 김홍식 (충북대학교 식물자원학과) ;
  • 우선희 (충북대학교 식물자원학과)
  • Received : 2014.12.14
  • Accepted : 2015.02.24
  • Published : 2015.03.31

Abstract

In the present study, different expression of protein from Taekwang was revealed by 2-DE, and expressions of protein on each week after flowering was investigated. After analysis of expression of protein, MALDI-TOF was executed to identify expected protein function. Results revealed that there were three patterns of expression of protein during the maturing. The first pattern was that proteins were gradually expressed as up-regulation from 1 week to 6 week. The second pattern was that proteins were expressed gradually from 1 week to 5 week and then it started down-regulation in 6 week. The last pattern was that proteins were gradually as up-regulation from 1 week to 3 week and then down-regulation until 6 week. This phenomenon suggests that young stage has more protein related to correspondence mechanism against disease and growth and then maturing stage has more expression of protein related to storage protein. In MALDI-TOF analysis, p24 oleosin isoform A protein was identified that relates oleosin which is synthetic product in oil body. This protein spot increased gradually until 5 week and then decreased after 5 week. It explained that the protein is active until maturing stage to protect oil in seed and then its activity has gradually degraded. This result may be expected that a protein, related to growth of a seed has increased until maturing and then a seed fills up with a storage protein.

본 연구는 국내 육성 품종인 태광콩의 등숙기에 따른 단백질 발현 양상을 비교함으로써 등숙기 단백질 발현의 차이에 대한 기초자료를 얻고자 수행하였다. 동한 개화 후 종실의 등숙이 진행됨에 따라서 단백질 발현 양상이 세가지 경향으로 나뉘어 지는 것을 확인하였다. 첫 번째는 등숙이 진행됨에 따라서 단백질 발현 정도가 증가하다가 감소되며, 두 번째는 증가와 감소의 시기가 성숙기에 이루어지며, 세번째는 등숙기부터 성숙기까지 점진적으로 증가하는 것이다. 이러한 현상은 단백질의 기능에 따라 달라지는 것으로 사료된다. 등숙 초기에는 등숙에 필요한 단백질의 발현이 증가할 것이며 등숙 후기에는 저장단백질의 발현이 증가할 것으로 사료된다. 따라서 향후 좀 더 많은 수의 단백질 spot 들을 동정하여 어떤 기능을 가진 단백질이 등숙기에 따라 단백질의 발현 양상이 달라지는지는 좀 더 면밀히 관찰할 필요성이 있다고 사료된다.

Keywords

References

  1. Bradford, M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Chem. 72 : 248-254.
  2. Gayler, K. R. and G. E. Sykes. 1981. ${\beta}$-Conglycinins in developing soybean seeds. Plant Physiol. 67 : 958-961. https://doi.org/10.1104/pp.67.5.958
  3. Hajduch, M., A. Ganapathy, J. W. Stein, and J. J. Thelen. 2005. A systematic proteomic study of seed filling in soybean. Plant Physiology. 137 : 1397-1419 https://doi.org/10.1104/pp.104.056614
  4. Hill, J. E. and R. W. Breidenbach. 1974. Proteins of soybean seeds. II. Accumulation of the major protein components during seed development and maturation. Plant Physiol 53 : 747-751. https://doi.org/10.1104/pp.53.5.747
  5. Katavic, V., G. K. Agrawal, M. Hajduch, S. L. Harris, and J. J. Thelen. 2006. Protein and lipid composition analysis of oil bodies from two Brassica napus cultivars. proteomics. 6 : 4586-4598. https://doi.org/10.1002/pmic.200600020
  6. Krishnan, H. B., G. Jiang, H. A. Krishnan, and J. W. Wiebold. 2000. Seed storage protein composition of non-nodulating soybean [Glycine max (L.) Merr.] and its influence on ptotein quality. Plant Sci. 157 : 191-199. https://doi.org/10.1016/S0168-9452(00)00283-1
  7. Krishnan, H. B. 2002. Evidence for accumulation of the ${\beta}$-subunit of ${\beta}$-conglycinin in soybean [Glycine max (L) Merr.] embryonic axes. Plant Cell Rep. 20 : 869-875. https://doi.org/10.1007/s00299-001-0400-5
  8. Loer, D. S. and Herman, E. M. 1993. Cotranslational integration of soybean (Glycine max) oil body membrane protein oleosin into microsomal membranes. Plant Physiol. 101 : 993-998.
  9. Mienke, D. W., J. Chen, and R. N. Beachy. 1981. Expression of storage-protein genes during soybean seed development. Planta. 153 : 130-139. https://doi.org/10.1007/BF00384094
  10. Nielsen, N. C. 1996. Soybean seed composition. In: Verma DPS, Shoemaker RC (eds) Soybean : genetics, molecular biology and biotechnology. CAB. Wallingford. 127-163.
  11. O'Farrell, P. F. 1975. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250 : 4007-4021.
  12. Rahman, Md. M., Md. M. Hossain, Md. P. Anwar, and A. S. Juraimi. 2011. Plant density influence on yield and nutritional quality of soybean seed. Asian J. of Plant Sci. 10 : 125-132. https://doi.org/10.3923/ajps.2011.125.132
  13. Roberts, R. C. and D. R. Briggs. 1965. Isolation and characterization of the 7S component of soybean globulins. Cereal Chem. 42 : 71-85.
  14. Thanh, V. H. and K. Shibasaki. 1976. Heterogeneity of betaconglycinin. Biochim Biophys Acta. 181 : 404-409.
  15. Woo, S. H., H. S. Kim, B. H. Song, C. W. Lee, Y. M. Park, S. K. Jong, and Y. G. Cho. 2003. Rice proteomics: A functional analysis of the rice genome and application. Reprintes from Korea J. of Plant Biotechnol. 30(3) : 281-291. https://doi.org/10.5010/JPB.2003.30.3.281