MALGORZATA OZGO, ADAM LEPCZYNSKI, AGNIESZKA HEROSIMCZYK

Two-dimensional gel-based serum protein profile of growing piglets

In the present study we have introduced a proteomic workflow, 2-dimensional electrophoresis coupled with matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) to create a 2-D map reflecting the characteristic pattern of serum proteins in growing piglets. Blood samples were collected from 6 piglets aged 50 days. Serum proteins were resolved at pH 3-10, followed by 12% SDS-PAGE. Reproducible protein spots (183) were picked from the gels and submitted for identification by peptide mass fingerprinting. As a result, 105 spots were positively identified, representing the products of 37 unique genes. Nearly half of the identified proteins (48.57%) were associated with transport and binding, 25.71% were classified as commonly circulating blood proteins, 11.43% as members of coagulation and complement factors, 8.57% as protease inhibitors, and 5.72% as proteases. We present 12 proteins that have not been previously identified in porcine serum/plasma samples using proteomic tools. Therefore, the gel-based map demonstrated in the current study complements currently available 2-D profiles of porcine plasma or serum and fills the gap between the existing protein patterns of neonatal and mature individuals. Moreover, the results of this work may be useful in monitoring changes in protein concentrations in young growing piglets in response to various physiological or pathological factors.

Anahtar Kelimeler:

Piglet, serum, proteome, 2-dimensional electrophoresis, MALDI-TOF MS

Two-dimensional gel-based serum protein profile of growing piglets

Keywords:

Piglet, serum, proteome, 2-dimensional electrophoresis, MALDI-TOF MS,

PDF

___

Ahn JY, Kim IY, Oh SJ, Hwang HS, Yi SS, Kim YN, Shin JH, Yoon YS, Seong JK (2014). Proteomic analysis of domestic pig pancreas during development using two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Lab Anim Res 30: 45–53.
Bendixen E, Danielsen M, Larsen K, Bendixen C (2010). Advances in porcine genomics and proteomics - a toolbox for developing the pig as a model organism for molecular biomedical research. Brief Funct Genomics 9: 208–219.
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.
Caperna TJ, Shannon AE, Garrett WM (2008). A gel-based reference map of the porcine hepatocyte proteome. Domest Anim Endocrinol 35: 142–156.
Chen X, Huang Z, Zhou B, Wang H, Jia G, Qiao J (2014). Expression and purification of porcine Akirin2 in Escherichia coli. Turk J Biol 38: 339–345.
de Almeida AM, Bendixen E (2012). Pig proteomics: a review of a species in the crossroad between biomedical and food sciences. J Proteomics 75: 4296–4314.
Ellederova Z, Halada P, Man P, Kubelka M, Motlik J, Kovarova H (2004). Protein patterns of pig oocytes during in vitro maturation. Biol Reprod 71: 1533–1539.
Esteso G, Mora MI, Garrido JJ, Corrales F, Moreno A (2008). Proteomic analysis of the porcine platelet proteome and alterations induced by thrombin activation. J Proteomics 71: 547–560.
Huang SY, Lin JH, Chen YH, Chuang CK, Lin EC, Huang MC, Sunny Sun HF, Lee WC (2005). A reference map and identification of porcine testis proteins using 2-DE and MS. Proteomics 5: 4205–4212.
Jeong JY, Nam JS, Kim JM, Jeong HJ, Kim KW, Lee HJ (2013a). Comparison of plasma proteome expression between the young and mature adult pigs. Reprod Dev Biol 37: 247–253.
Jeong JY, Nam JS, Park MR, Kim JM, Jeong HJ, Kim KW, Lee HJ (2013b). A proteome reference map for porcine plasma proteins. Reprod Dev Biol 37: 255–261.
Lahm HW, Langen H (2000). Mass spectrometry: a tool for the identification of proteins separated by gels. Electrophoresis 21: 2105–2114.
Miller I, Wait R, Sipos W, Gemeiner M (2009). A proteomic reference map for pig serum proteins as a prerequisite for diagnostic applications. Res Vet Sci 86: 362–367.
Pérez-Reinado E, Ramírez-Boo M, Garrido JJ, Jorrín JV, Moreno A (2007). Towards a global analysis of porcine alveolar macrophages proteins through two-dimensional electrophoresis and mass spectrometry. Dev Comp Immunol 31: 1220–1232.
Verma N, Rettenmeier AW, Schmitz-Spanke S (2011). Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies. Proteomics 11: 776–793.
Wang H, Chen X, Huang Z, Zhou B, Jia G, Liu G, Zhao H (2014). Expression and purification of porcine PID1 gene in Escherichia coli. Turk J Biol 38: 523–527.
Westermeier R (2006). Sensitive, quantitative and fast modifications for Coomassie Blue staining of polyacrylamide gels. Proteomics 6 (Suppl. 1): 61–64.
Zhang K, Liu Y, Shang Y, Zheng H, Guo J, Tian H, Jin Y, He J, Liu X (2012). Analysis of pig serum proteins based on shotgun liquid chromatography-tandem mass spectrometry. Afr J Biotechnol 11: 12118–12127.