STUDY ON PROTEOMICS EXPRESSION PROFILE OF SERA FROM PATIENTS WITH UREMIA

Yıl 2008, Cilt: 5 Sayı: 2, 64 - 73, 01.06.2008

Öz

-

Anahtar Kelimeler

-

STUDY ON PROTEOMICS EXPRESSION PROFILE OF SERA FROM PATIENTS WITH UREMIA

Öz

Aim: To establish and optimize two-dimensional electrophoresis (2DE) and relevant techniques for the study of serum proteome of the patients with uremia, and compare serum protein 2DE pattern between the uremia patients and the normal. Methods: Using immobiline pH gradients isoelectricfocusing (IEF) as the 1st dimension and vertical SDS-PAGE as the 2nd dimension, we performed the comparison and conditional optimization of the factors. Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis,silver staining, ImageMaster 2D 5.0 analysis software, matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS) and IPI human database searching, were used to separate and indentify the proteome of the sera from the patients with uremia. Results: Satisfactory 2DE patterns of the serum proteins were obtained. 26 protein spots were remarkably hanged in uremia patients, and 20 protein spots were identified by MALDI-TOF-TOFMS.. Conclusion: Good reproducibility could be obtained by applying immobilized pH gradient 2DE to separate and identify the proteome in sera, which contributes to the further study on uremia toxins pertaining to protein

Anahtar Kelimeler

Uremia, proteome, MALDI-TOF-TOF-MS, two-dimensional gel electrophoresis

Kaynakça

• Hou Fan-fan. New knowledge of uremia toxins. Chinese J Nephrology 2003;19:69-70
• Guo Yao-jun. Experimental Techniques of Protein Electrophoresis. Beijing: Science Press,1999, 141-2
• Gorg A,Postel W,Gunther S. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 2000;21:1037-53
• Sanchez JC,Rouge V,Pisteur M,et al.Improved and simplified in-gel sample application using reswelling of dry immobilized pH gradients. Electrophoresis 1997;18:324-7
• Gharahdaghi F, Weinberg C, Meagher D, et al. Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity. Electrophoresis 1999;20:601-5
• Tu Cheng-jian. Holographic Preparation of Sample. Referred to Xia Qi-chang, Protein Chemistry and Proteome. Beijing: Science Press, 2004, 263-4
• Georgiou HM, Rice GE,Baker MS.Proteomic analysis of human plasma:failure of entrifugal ultrafiltration to remove albumin and other high molecular weigh proteins. Proteomics 2001;1(12):1503-6
• Liu WJ, Shen Y, Ding J. Protein phosphatase 2A: Its structure, function and activity regulation. Acta Biochimica Polonica 2001;44:921-30
• Kopple JD, Mehrotra R, Suppasyndh O, et al. Observations with regard to the National Kidney Foundation K/DOQI clinical practice guidelines concerning serum transthyretin in chronic renal failure. Clin Chem Lab Med 2002;40: 1308-12
• Monica MS, Anthony GW, Norden, et al. Evidence for the role of megalin in renal uptake of transthyretin. Biol Chem 2000;275: 176-181
• Behren TW, Jagadeesh J, Scherle P, et al. Jawl a lymphoid restricted membrane protein localized to the endoplasmic reticulum. J Immunol 1994;153(2):682-90
• Sukotjo C, Abanmy AA, Ogawa T. Molecular cloning of wound inducible transcript (wit 3.0) differentially expressed in edentulous oral mucosa undergoing tooth extraction wound-healing. J Dent Res 2002;81:229-35
• Thomas J G, Ayling A, Baneyx F. Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins fom E. Coli: to fold or refold. Appl Biochem Biotechnol 1997;66:41-2
• Michele R W, David M B, HardenT K. PLC-ɛ: A shared effector protein in Ras-, Rho-, and Gαβγ-mediated signaling. Mol Interventions 2003;3:273-80
• Thiede B, Dimmler C, Siejak F. Predominant identification of RNA-binding proteins in Fas induced apoptosis by proteome analysis. J Biol Chem 2001;276: 44-50