血清中低分子量蛋白质的提取和鉴定

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (10): 1955.

血清中低分子量蛋白质的提取和鉴定

李铁纯1,2, 汪子明1, 王璐1, 张寒琦1, 于永3

1. 吉林大学化学学院, 长春 130012;
2. 鞍山师范学院化学系, 鞍山 114007;
3. 吉林大学仪器科学与电气工程学院，长春 130061

收稿日期:2010-03-08 出版日期:2010-10-10 发布日期:2010-10-10
通讯作者: 于永, 男, 博士, 讲师, 主要从事分析仪器与分析方法研究. E-mail: yuyong@jlu.edu.cn

Extraction and Identification of the Low Molecular Weight Proteins in Serum

LI Tie-Chun1,2, WANG Zi-Ming1, WANG Lu1, ZHANG Han-Qi1, YU Yong3*

1. College of Chemistry, Jilin University, Changchun 130012, China;
2. Department of Chemistry, Anshan Normal University, Anshan 114007, China;
3. College of Instrumentation and Electrical Engineering, Jilin University, Changchun 130061, China

Received:2010-03-08 Online:2010-10-10 Published:2010-10-10
Contact: YU Yong. E-mail: yuyong@jlu.edu.cn

摘要/Abstract

摘要： 采用改进的圆盘凝胶电泳提取人血清中低分子量蛋白质, 去除了血清中分子量大于3×104的蛋白质, 将提取的低分子量蛋白质热变性后直接在溶液中酶解成肽, 经液相色谱-质谱分析, 并进行Mascot数据库检索, 确认出人血清中97种蛋白质.

关键词: 血清, 低分子量蛋白, 液相色谱串联质谱, 圆盘凝胶电泳

Abstract: An improved disc sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDS-PAGE) to extract low molecular weight(LMW) proteins from human and cutoff proteins larger than 3×104 was developed. In the disc SDS-PAGE, short column resolving gel and little of electrode buffer solutions were used in mini home-made electrophoresis equipment. Passive elution of the LMW proteins from the gel was adopted without staining. After removing SDS and precipitation, the obtained LMW protein mixtures were denatured directly in solution and degraded in the presence of trypsin to obtain peptides. The peptides were analyzed by HPLC/MS/MS. The LMW proteins were identified by matching the peptide patterns in mass spectra with those in Mascot data pool and 97 kinds of LMW proteins in human serum were identified.

Key words: Serum, Low molecular weight protein, HPLC/MS/MS, Disc PAGE

TrendMD:

李铁纯, 汪子明, 王璐, 张寒琦, 于永. 血清中低分子量蛋白质的提取和鉴定. 高等学校化学学报, 2010, 31(10): 1955.

LI Tie-Chun, WANG Zi-Ming, WANG Lu, ZHANG Han-Qi, YU Yong*. Extraction and Identification of the Low Molecular Weight Proteins in Serum. Chem. J. Chinese Universities, 2010, 31(10): 1955.

参考文献

[1]Anderson N. L., Anderson N. G.. Mol. Cell Proteomics[J], 2002, 1: 845—867 [2]Tirumalai R. S., Chan K. C., Prieto D. A., Issaq H. J., Conrads T. P., Veenstra T. D.. Mol. Cell Proteomics[J], 2003, 2: 1096—1103 [3]Bischoff R., Luider T. M.. J. Chromatogr. B[J], 2004, 803: 27—40 [4]Wittke S., Fliser D., Haubitz M., Bartel S., Krebs R., Hausadel F., Hillmann M.. J. Chromatogr. A[J], 2003, 1013: 173—181 [5]Dean C., Jesse G., Paul T.. J. Gen. Intern. Med.[J], 2008, 23: 78—84 [6]Qiu J. G., Fan J., Liu Y. K., Zhou J., Dai Z., Huang C., Tang Z. Y.. J. Cancer Res. Clin. Oncol.[J], 2008, 134: 299—305 [7]Petricoin E. F., Belluco C., Araujo R. P., Liotta L. A.. Nat. Rev. Cancer[J], 2006, 6(12): 961—967 [8]Callesen A. K., Mohammed S., Bunkenborg J., Kruse T. A., Cold S., Mogensen O., Kruse T. A., Jensen O. N., Madsen J. S.. Rapid Commun. Mass Spectrom.[J], 2005, 19: 1578—1586 [9]Chernokalskaya E., Gutierrez S., Pitt A. M., Leonard J. T.. Electrophoresis[J], 2004, 25: 2461—2468 [10]Berven F. S., Kroksveen A. C., Berle M., Rajalahti T., Flikka K., Arneberg R., Myhr K. M., Vedeler C., Kvalheim O. M., Ulvik R.. J. Proteomics Clinical Applications[J], 2007, 7: 699—711 [11]Aresta A., Calvano C. D., Zambonin C. G., Monaco A., Tommasi S., Pilato B., Paradiso A.. J. Pharm. Biomed. Anal.[J], 2008, 46: 157—164 [12]Zheng X. Y., Baker H., Hancock W. S.. J. Chromatogr. A[J], 2006, 1120: 173—184 [13]Harper R. G., Workman S. R., Schuetzner S., Timperman A. T., Sutton J. N.. Electrophoresis[J], 2004, 25: 1299—1306 [14]Huang H. L., Stasyk T., Morandell S., Mogg M., Schreiber M., Feuerstein I., Huck C. W., Stecher G., Bonn G. K., Huber L. A.. Electrophoresis[J], 2005, 26: 2843—2849 [15]Liu X. L., Li L. J., Zhang G. Y., Sheng G. P., Xu W.. Clin. Biochem.[J], 2007, 40: 1266—1271 [16]Li T. C., Dai S. P., Wang Z. M., Zhang H. Q.. Electrophoresis[J], 2010, 31: 1090—1096 [17]Schagger H., Jagow G. V.. Anal. Biochem.[J], 1987, 166: 368—379 [18]Klein C., Aivaliotis M., Olsen J. V., Falb M., Besir H., Scheffer B., Bisle B., Falb M., Pfeiffer F., Siedler F., Oesterhelt D.. J. Proteome Res.[J], 2007, 4: 1510—1518 [19]Park Z. Y., Russell D. H.. Anal. Chem.[J], 2001, 73: 2558—2564 [20]Adkins J. N., Varnum S. M., Auberry K. J., Angell N. H., Smith R. D., Springer D. L., Pounds J. G.. Mol. Cell Proteomics[J], 2002, 1(12): 947—955