Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
권호 표지
DOI QR코드

DOI QR Code

Comparative Analysis of Muscle Proteome from Porcine White and Red Muscles by Two-dimensional Electrophoresis

이차원전기영동법을 이용한 white muscle과 red muscle간의 단백질 발현양상의 비교분석

  • Kim, N.K. (Department of Applied Biochemistry) ;
  • Joh, J.H. (Department of Applied Biochemistry) ;
  • Chu, K.S. (Department of Applied Biochemistry) ;
  • Park, H.R. (Department of Applied Biochemistry) ;
  • Park, B.Y. (National Livestock Research Institute, R.D.A.) ;
  • Kim, O.H. (Department of Animal Science, College of Natural Science, Konkuk University) ;
  • Lee, C.S. (Department of Applied Biochemistry)
  • 김남국 (건국대학교 자연과학대학 응용생화학) ;
  • 조중호 (건국대학교 자연과학대학 응용생화학) ;
  • 추교선 (건국대학교 자연과학대학 응용생화학) ;
  • 박혜란 (건국대학교 자연과학대학 응용생화학) ;
  • 박범영 (농촌진흥청 축산기술연구소) ;
  • 김언현 (건국대학교 자연과학대학 축산학과) ;
  • 이창수 (건국대학교 자연과학대학 응용생화학)
  • Published : 2003.10.31
Download PDF
⟨ Previous Next ⟩

Abstract

The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

근육의 생화학적 특성을 단백질 수준에서 분석하기 위하여 3원교잡종 돼지 3개체를 선발하고, white muscle은 longissimus dorsi muscle을 red muscle은 soleus muslce을 분리하여 분석에 이용하였다. 각 근육조직은 수용성, 불수용성 단백질 및 총단백질로 분리하여 추출하였고, 이차원전기영동 분석을 위하여 17cm 길이의 immobilized pH gradient strip (Bio-Rad, 3-10NL)과 12% acrylamide gel을 이용하여 전개하였다. 각각의 gel은 coomassie stain과 silver stain을 통하여 가시화 하였고, PDQuest software을 통하여 단백질 발현양상을 분석하였다. 하나의 gel에서 평균 600개 이상의 단백질 spot을 관찰하였으며, 반복실험을 통하여 white muscle과 red muscle간에 발현의 차이를 보이는 5개의 단백질 spot을 확인할 수 있었다. 5개의 spot 중 4개의 단백질은 측정된 분자량과 pI값이 troponin I, T 및 myoglobin의 수치값과 유사한 것으로 확인되었다. 그러나, 1개의 spot은 오차범위 내에서 유사한 단백질을 확인할 수 없었다. 5개의 단백질 spot중 1개(spot 1)는 white muscle에서, 4개의 spot(spot 2～5)은 red muscle에서 높게 발현됨을 확인하였으며, 특히 spot 4의 경우 white muscle 보다 red muscle에서 평균 14.6배 높게 발현됨을 확인하였다. 본 연구는 근육의 생화학적 특성을 이해하는데 중요한 기초 자료로 활용될 수 있으며, 앞으로 white muscle과 red muscle의 단백질 발현 분석을 통하여 단백질 수준에서의 생화학적 특성에 관한 연구가 충분히 진행되어야 할 것이다.

Keywords

References

  1. Beecher, G. R., Kastenschmidt, L. L., Cassens, R. G., Hoekstra, W. G. and Briskey, E. J. 1968. A comparison of the light and dark portions of a striated muscle. J. Food Sci. 33:84.
  2. Cassens, R. G. and Cooper, C. C. 1971. Red and white muscle. Adv. Food Res. 19:1.
  3. Cornet, M. and Bousset, J. 1999. Free amino acids and dipeptides in procine muscles: differences between red and white muscles. Meat Sci. 51:215.
  4. Essen-Gustavsson, B., Karlstrom, K. and Lundstrom, K. 1992. Muscle fibre characteristics and metabolic response at slaughter in pigs of different halothane genotypes and their relation to meat quality. Meat Sci. 31:1.
  5. Fernandez, X., Lefaucheur, L. and Candek, M. 1995. Comparative study of two classifications of muscle fibres:consequences for the photometric determination of glycogen according to fibre type in red and white muscle of the pig. Meat Sci. 41:225.
  6. Henckel, P., Oksbjerg, N., Erlandsen, E., Barton-Gade, P. and Bejerholm, C. 1997. Histo- and biochemical characteristics of the longissimus dorsi muscle in pigs and their relationships to performance and meat quality. Meat Sci. 47:311.
  7. Karlsson, A., Enfalt, A. C., Essen-Gustavsson, B., Lundstrom, K., Rydhmer, L. and Stern, S. 1993. Muscle histochemical and biochemical properties in relation to meat quality during selection for increased lean tissue growth rate in pigs. J. Anim. Sci. 71:930.
  8. Karlsson, A. H., Klont, R. E. and Fernandez, X. 1999. Skeletal muscle fibres as factors for pork quality. Livest. Prod. Sci. 60:255.
  9. Klont, R. E., Brocks, L. and Eikelenboom. G. 1998. Muscle fibre type and meat quality. Meat Sci. 49:S219.
  10. Lawrie, R. A. 1950. Some observations on factors affecting myoglobin concentrations in muscle. J. Agr. Sci. 40:356.
  11. Lawrie, R. A. 1985. Meat science, 4th ed. Pergamon press, Headington Hill Hall, Oxford, U.K., p43.
  12. Maltin, C. A., Warkup, C. C., Matthews, K. R., Grant, C. M., Porter, A. D. and Delday, M. I. 1997. Pig muscle fibre characteristics as a source of variation in eating quality. Meat Sci. 47:237.
  13. Margreth, A. and Novello, F. 1964. Observations on the chemical determination and distribution of ribonucleic acid in several striated muscle. Exp. Cell Res. 35:38.
  14. Molloy, M. P., Herbert, B. R., Walsh, B. J., Tyler, M. I., Traini, M., Sanchez, J. C., Hochstrasser, D. F., Williams, K. L. and Gooley, A. A. 1998. Extraction of membrane proteins by differential solubilization for separation using two-dimensional gel electrophoresis. Electrophoresis. 19:837.
  15. Neuhoff, V., Arold, N., Taube, D. and Ehrhardt, W. 1988. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using coomassie brilliant blue G-250 and R-250. Electrophoresis. 9:255.
  16. Ozawa, S., Mitsuhashi, T., Mitsumoto, M., Matsumoto, S., Ttoh, N., Itagaki, K., Kohno, Y. and Dohgo, T. 2000. The characteristics of muscle fiber types of longissimus thoracis muscle and their influences on the quantity and quality of meat from japanese black steers. Meat Sci. 54:65.
  17. Pandey, A. and Mann, M. 2000. Proteomics to study genes and genomes. Nature 405:837.
  18. Pette, D. and Staron, R. S. 1990. Cellular and molecular diversities of mammalian skeletal muscle fibres. Rev. Physiol. Biochem. Pharmacol. 116:1.
  19. Shevchenko, A., Wilm, M., Vorm, O. and Mann, M. 1996. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68:850.
  20. Soren, N. H., Michael, D. W. and Rainer Cramer. 2001. Proteomics-post-genomic cartography to understand gene function. Pharmacol. Sci. 22:376.
  21. Takasu, T. and Hunhes, B. P. 1966. Lactate dehydrogenase isoenzymes in developing human muscle. Nature 212:609.