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Buffer optimization for bovine Longissimus muscle tissues: Proteome analysis of Korean native cattle using 2-dimensional gel electrophoresis

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Because beef contains a high proportion of marbling, we sought to optimize the solubilization of the longissimus muscle proteome using modified lysis and isoelectric focusing (IEF) rehydration buffers. Of the 3 lysis buffers tested, those containing a thiourea-urea mixture provided superior resolution, whereas that which contained only urea yielded consistently poor results. In addition, we found that the IEF rehydration buffer containing a thiourea-urea mixture, a high dithiothreitol level, and Pharmalyte was able to generate sharper 2-dimensional (2-D) gels, whereas that which contained immobilized pH gradient (IPG) buffer had no an effect on protein resolution. The number of different protein spots between the muscle-development stage (11 months) and the fat-development stage (17 months) in Hanwoo steers was 39, whereas the use of normal buffer resulted in a detection of a difference of only 7 protein spots. This study provides a methodological tool for studying the proteome of bovine longissimus muscle with a high fat content using 2-D gel electrophoresis (2-DE) gels.

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  1. Shackelford SD, Wheeler TL, Koohmaraie M. Relationship between shears force and trained sensory panel tenderness ratings of 10 major muscles from Bos indicus and Bos Taurus cattle. J. Anim. Sci. 73: 3333–3340 (1995)

    CAS  Google Scholar 

  2. Harper GS, Pethick D. The physiology of marbling: What is it, and why does it develop? pp. 34–45. In: Marbling Symposium. October 3, Coffs Harbour, Cooperative Research Centre for Cattle and Beef Quality. Armidole, NSW, Australia (2001)

    Google Scholar 

  3. Santoni V, Kieffer S, Desclaux D, Masson F, Rabilloud T. Towards the recovery of hydrophobic proteins on two-dimensional electrophoresis gels. Electrophoresis 21: 3329–3344 (2000)

    Article  CAS  Google Scholar 

  4. Wang SB, Hu Q, Sommerfeld M, Chen F. An optimized protocol for isolation of soluble proteins from microalgae for two-dimensional gel electrophoresis analysis. J. Appl. Phycol. 15: 485–496 (2003)

    Article  CAS  Google Scholar 

  5. Ruan Y, Wan M. An optimized procedure for solubilization, reduction, and transfer of human breast cancer membrane-enriched fraction by 2-DE. Electrophoresis 28: 3333–3340 (2007)

    Article  CAS  Google Scholar 

  6. Rabilloud T. Solubilization of proteins for electrophoretic analyses. Electrophoresis 17: 813–829 (1996)

    Article  CAS  Google Scholar 

  7. Rabilloud T, Adessi C, Giraudel A, Lunardi J. Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 18: 307–316 (1997)

    Article  CAS  Google Scholar 

  8. Cheah KS, Cheah AM, Just A. Simple and rapid biopsy technique for obtaining fat and muscle samples from live animals for predicting meat quality. Dan. Veterinaertidsskr. 80: 775–777 (1997)

    Google Scholar 

  9. Gelfi C, De Palma S, Cerretelli P, Begum S, Wait R. Twodimensional protein map of human vastus lateralis muscle. Electrophoresis 24: 286–295 (2003)

    Article  CAS  Google Scholar 

  10. Musante L, Candiano G, Ghiggeri GM. Resolution of fibronectin and other uncharacterized proteins by two-dimensional polyacrylamide electrophoresis with thiourea. J. Chromatogr. B 13: 351–356 (1998)

    Google Scholar 

  11. Molloy MP, Herbert BR, Walsh BI, Tyler MI, Traini M, Sanchez JC, Hochstrasser DF, Williams KL, Gooley AA. Extraction of membrane proteins by differential solubilization for separation using two-dimensional gel electrophoresis. Electrophoresis 19: 837–844 (1998)

    Article  CAS  Google Scholar 

  12. Görg A, Obermaier C, Boguth G, Harder A, Scheibe B, Wildgruber R, Weiss W. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 21: 1037–1053 (2000)

    Article  Google Scholar 

  13. Majercakova P, Kucerova Z, Desvaux FX, Peltre G. Electrotitration curves of human gastric pepsinogens in agarose gels. Electrophoresis 21: 2919–2924 (2000)

    Article  CAS  Google Scholar 

  14. Busnel JM, Kilar F, Kasicka V, Descroix S, Hennion MC, Peltre G. Carrier ampholytes-based capillary electrophoresis as an alternative to capillary zone electrophoresis in classical background electrolytes. J. Chromatogr. A 16: 183–188, 1087 (2005)

    Article  Google Scholar 

  15. Stedry M, Jaros M, Vcelakova K, Gas B. Eigenmobilities in background electrolytes for capillary zone electrophoresis: II. Eigenpeaks in univalent weak electrolytes. Electrophoresis 24: 536–547 (2003)

    Article  CAS  Google Scholar 

  16. Chen AB, Rickel CA, Flanigan A, Hunt G, Moorhouse KG. Comparison of ampholytes used for slab gel and capillary isoelectric focusing of recombinant tissue-type plasminogen activator glycoforms. J. Chromatogr. A 13: 279–284, 744 (1996)

    Article  CAS  Google Scholar 

  17. Herbert BR, Molloy MP, Gooley AA, Walsh BJ, Bryson WG, Williams KL. Improved protein solubility in two-dimensional electrophoresis using tributyl phosphine as reducing agent. Electrophoresis 19: 845–851 (1998)

    Article  CAS  Google Scholar 

  18. Berkelman T, Stenstedt T. 2-D Electrophoresis Using Immobilized pH Gradients, Principles, and Methods. 2nd ed. Method 80-6429-60. Amersham Biosciences, Uppsala, Sweden (2002)

    Google Scholar 

  19. Talamo F, D’Ambrosio C, Arena S, Vecchio PD, Ledda L, Zehender G, Ferrara L, Scaloni A. Proteins from biwine tissues and biological fluids: Defining a reference electrophoresis map for liver, kindny, muscle, plasma, and red blood cells. Proteomics 3: 440–460 (2003)

    Article  CAS  Google Scholar 

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Correspondence to Yunjaie Choi.

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Lee, H., Han, J., Lee, K. et al. Buffer optimization for bovine Longissimus muscle tissues: Proteome analysis of Korean native cattle using 2-dimensional gel electrophoresis. Food Sci Biotechnol 19, 1107–1112 (2010).

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