Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

  • Han, Rong-Xun (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Choi, Su-Min (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Kim, Myung-Youn (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Quan, Yan Shi (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Kim, Baek-Chul (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Diao, Yun Fei (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Koqani, Reza (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Park, Chang-Sik (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University) ;
  • Jin, Dong-Il (Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs Chungnam National University)
  • 발행 : 2008.12.31

초록

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

키워드

참고문헌

  1. Ali A, Coenen K, Bousquet D, Sirard MA (2004): Origin of bovine follicular fluid and its effect during in vitro maturation on the developmental competence of bovine oocytes. Theriogenology 62:1596-1606 https://doi.org/10.1016/j.theriogenology.2004.03.011
  2. Arima Y, Bremel RD (1983): Purification and characterization of bovine placental lactogen. Endocrinology 113:2186-2194 https://doi.org/10.1210/endo-113-6-2186
  3. Bertram Y Fong, Carmen S Norris, Kate P Palmano (2008): Fractionation of bovine whey proteins and characterisation by proteomic techniques. Int Dairy J 18:23-46 https://doi.org/10.1016/j.idairyj.2007.06.005
  4. Bouley J, Chambon C, Picard B (2004): Mapping of bovine skeletal muscle proteins using two-dimensional gel electrophoresis and mass spectrometry. Proteomics 4:1811-1824 https://doi.org/10.1002/pmic.200300688
  5. Farrell HM Jr, Jimenez-Flores R, Bleck GT, Brown EM, Butler JE, Creamer LK, Hicks CL, Hollar CM, Ng-Kwai-Hang KF, Swaisgood HE (2004): Nomenclature of the proteins of cows' milk-sixth revision. J Dairy Sci 87:1641-1674 https://doi.org/10.3168/jds.S0022-0302(04)73319-6
  6. Galvani M, Hamdan M, Righetti PG (2001): Two-dimensional gel electrophoresis/matrix-assisted laser desorption/ionisation mass spectrometry of commercial bovine milk. Rapid Commun Mass Spectrom 15:258-264 https://doi.org/10.1002/rcm.220
  7. Gorg A, Obermaier C, Boguth G, Harder A, Scheibe B, Wildgruber R, Weiss W (2000): The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 21:1037-1053 https://doi.org/10.1002/(SICI)1522-2683(20000401)21:6<1037::AID-ELPS1037>3.0.CO;2-V
  8. Hurley WL (1989): Mammary gland function during involution. J Dairy Sci 72:1637-1646 https://doi.org/10.3168/jds.S0022-0302(89)79276-6
  9. Jalili PR, Gheyi T, Dass C (2004): Proteome analysis in bovine adrenal medulla using matrix-assisted laser desorption/ionization mass spectrometry. Rapid Commun Mass Spectrom 18:914-916 https://doi.org/10.1002/rcm.1422
  10. Jobim MI, Oberst ER, Salbego CG, Souza DO, Wald VB, Tramontina F, Mattos RC (2004): Two-dimensional polyacrylamide gel electrophoresis of bovine seminal plasma proteins and their relation with semen freezability. Theriogenology 15:255-266
  11. Kim HR, Kang JK, Lee HR, Yoon JT, Seong HH, Jung JK, Park CS, Jin DI (2005): Cloned placenta of korean native calves died suddenly at two months after birth displays differential protein expression. Reprod Dev Biol 31:63-68
  12. Korhonen H, Marnila P, Gill HS (2000): Milk immunoglobulins and complement factors. Br J Nutr 84:75-80
  13. Lacy-Hulbert SJ, Woolford MW, Nicholas GD, Prosser CG, Stelwagen K (1999): Effect of milking frequency and pasture intake on milk yield and composition of late lactation cows. J Dairy Sci 82:1232-1239 https://doi.org/10.3168/jds.S0022-0302(99)75346-4
  14. Legrand D, Pierce A, Elass E, Carpentier M, Mariller C, Mazurier J (2008): Lactoferrin structure and functions. Adv Exp Med Biol 606:163-194 https://doi.org/10.1007/978-0-387-74087-4_6
  15. Lindmark-Mansson H, Timgren A, Aden G, Paulsson M (2005): Two-dimensional gel electrophoresis of proteins and peptides in bovine milk. Int Dairy J 15:111-121 https://doi.org/10.1016/j.idairyj.2004.06.010
  16. Lippolis JD, Reinhardt TA (2005): Proteomic survey of bovine neutrophils. Vet Immunol Immunopathol 103:53-65 https://doi.org/10.1016/j.vetimm.2004.08.019
  17. Lonnerdal Bo (1985): Biochemistry and physiological function of human milk proteins. Am J Clin Nutr 42:1299-1317 https://doi.org/10.1093/ajcn/42.6.1299
  18. Manso MA, Leonil J, Jan G, Gagnaire V (2005): Application of proteomics to the characterisation of milk and dairy products. Int Dairy J 15:845-855 https://doi.org/10.1016/j.idairyj.2004.07.021
  19. Radtke AL, O'Riordan MX (2008): Homeostatic maintenance of pathogen-containing vacuoles requires TBK1-dependent regulation of aquaporin-1. Cell Microbiol 10:2197-2207 https://doi.org/10.1111/j.1462-5822.2008.01199.x
  20. Sasser RG, Crock J, Ruder-Montgomery CA (1989): Characteristics of pregnancy-specific protein B in cattle. J Reprod Fertil Suppl 37:109-113
  21. Schuler LA, Shimomura K, Kessler MA, Zieler CG, Bremel RD (1988): Bovine placental lactogen: molecular cloning and protein structure. Biochemistry 27:8443-8448 https://doi.org/10.1021/bi00422a022
  22. Scott P, Kessler MA, Schuler LA (1992): Molecular cloning of the bovine prolactin receptor and distribution of prolactin and growth hormone receptor transcripts in fetal and utero-placental tissues. Mol Cell Endocrinol 89:47-58 https://doi.org/10.1016/0303-7207(92)90210-W
  23. Se'verin S, Wenshui X (2005): Milk biologically active components as nutraceuticals: Review. Crit Rev Food Sci Nutr 45:645-656 https://doi.org/10.1080/10408690490911756
  24. Smolenski G, Haines S, Kwan FY, Bond J, Farr V, Davis SR, Stelwagen K, Wheeler TT (2007): Characterisation of host defence proteins in milk using a proteomic approach. J Proteome Res 6:207-215 https://doi.org/10.1021/pr0603405
  25. Talamo F, D'Ambrosio C, Arena S, Del Vecchio P, Ledda L, Zehender G, Ferrara L, Scaloni A (2003): Proteins from bovine tissues and biological fluids: Defining a reference electrophoresis map for liver, kidney, muscle, plasma and red blood cells. Proteomics 3:440-460 https://doi.org/10.1002/pmic.200390059
  26. Wait R, Miller I, Eberini I, Cairoli F, Veronesi C, Battocchio M, Gemeiner M, Gianazza E (2002): Strategies for proteomics with incompletely characterized genomes: the proteome of Bos taurus serum. Electrophoresis 23:3418-3427 https://doi.org/10.1002/1522-2683(200210)23:19<3418::AID-ELPS3418>3.0.CO;2-7
  27. Ward PP, Uribe-Luna S, Conneely OM (2002): Lactoferrin and host defense. Biochem Cell Biol 80:95-102 https://doi.org/10.1139/o01-214
  28. Woo JH, Ko YG, Kim BK, Kim JM, Lee YS, Kim NY, Im GS, Yang BC, Seong HH, Jung JK, Kwun MS, Chung HJ (2005): Characterization of placental proteins in bovine somatic cell clone fetuses. Reprod Dev Biol 31:83-91
  29. Yamada M, Murakami K, Wallingford JC, Yuki Y (2002): Identification of low-abundance proteins of bovine colostral and mature milk using two-dimensional electrophoresis followed by microsequencing and mass spectrometry. Electrophoresis 23:1153-1160 https://doi.org/10.1002/1522-2683(200204)23:7/8<1153::AID-ELPS1153>3.0.CO;2-Y
  30. Zieler CG, Kessler MA, Schuler LA (1990): Characterization of a novel prolactin-related protein from bovine fetal placenta. Endocrinology 126:2377-2382 https://doi.org/10.1210/endo-126-5-2377