Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis

Chem. J. Chinese Universities ›› 2010, Vol. 31 ›› Issue (3): 502.

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Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis

SONG Jian^1,2, XU Jun-Jie^3,4, WEI Jing-Yan³, YU Yang³, SUN Wei-Guo², ZHANG Gui-Rong^2*

1. College of Electronic Science and Engineering,
2. College of Fundamental Medicine,
3. College of Pharmaceutical Science; Jilin University, Changchun 130021, China;
4. Department of Biochemistry, School of Basic Medical Sciences, Jilin Medicine College, Jilin 132013, China

Received:2009-05-14 Online:2010-03-10 Published:2010-03-10
Contact: ZHANG Gui-Rong. E-mail: zhanggr@jlu.edu.cn
Supported by:
国家自然科学基金(批准号: 30870540)和吉林省科技发展计划项目(批准号: 20070726, 20070425, 200705368)资助.

Abstract

Abstract:

Ala180 in site 1 and Ala44 in site 2 of human selenium-containing single chain abzyme Se-scFv-B3 were chosen to be mutated to serines, respectively, to study the substrate binding sites and the catalytic group of Se-scFv-B3 and to find the reason why the Se-scFv-B3 has lower catalytic activity based on the theoretical anticipation for the protein. After serine residue was reacted into secystins by chemical modification, the activity of the anterior one is about two times than that of the Se-scFv-B3, but the latter one did not offer a significant improvement. This indicated that site 1 was the predominant binding site for GSH, which was consistent with the foregoing conjecture and the results of energy calculation. In this study, the GPX activity of Se-scFv-B3 was improved and the site 1 preliminarily confirmed to be the substrate binding site based on both theoretical and experimental research. All of these observations will be useful for further work in this area.

Key words: Quikchange site-directed mutagenesis; Human selenium-containing sigle chain abzyme; Substrate-binding site

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SONG Jian, XU Jun-Jie, WEI Jing-Yan, YU Yang, SUN Wei-Guo, ZHANG Gui-Rong. Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis[J]. Chem. J. Chinese Universities, 2010, 31(3): 502.

References

[1]Müller A., Cadenas E., Graf P., et al... Biochem. Pharmacol.[J], 1984, 33: 3235—3239
[2]Back T. G., Dyck B. P.. J. Am. Chem. Soc.[J], 1997, 119: 2079—2083
[3]Liu J. Q., Gao S. J., Luo G. M., et al.. Biochem. Biophys. Res. Commol./Lun.[J], 1998, 247: 397—400
[4]Liu J. Q., Luo G. M., Ren X. J., et al.. Biochim. Biophys. Acta[J], 2000, 1481: 222—228
[5]Pauling L.. Am. Sci.[J], 1948, 36: 51—58
[6]Su D., Ren X. J., You D. L., et al.. Arch. Biochem. Biophys.[J], 2001, 395: 177 —184
[7]Luo G. M., Zhu Z. Q., Ding L., et al.. Biochem. Biophys. Res. Commol./Lun.[J], 1994, 198: 1240—1247
[8]Ding L., Liu Z., Zhu Z. Q., et al.. Biochem. J.[J], 1998, 332: 251—255
[9]Ren X. J., Gao S. J., You D. L., et al.. Biochem. J.[J], 2001, 359: 369—374
[10]Seccamani E., Tattanelli M., Mariani M., et al.. Int. J. Rad. Appl. Instrum. B.[J], 1989, 16: 167—170
[11]Reynolds J.C., DelVecchio S., Sakahara H., et al.. Nucl. Med. Biol.[J], 1989, 16: 121—125
[12]Huo R., Wei J. Y., Xu J. J., et al.. J. Mol. Recognit.[J], 2008, 21: 323—328
[13]Xu Jun-jie, Song Jian, Yan Fei, et al.. J. Mol. Recognit.[J], 2009, 22(44): 293—300
[14]HUO Rui(霍锐), SHI Yi(石毅), WEI Jing-Yan(魏景艳), et al.. Chem. J. Chinese Universities (高等学校化学学报)[J], 2008, 29(7): 1379—1383
[15]ZHANG Hao(张浩). Chin. J. Mol. Immunol.(免疫学杂志)[J], 2000, 16(4): 108—110
[16]Jiang J. H., Xiao J. Z., Yang S., et al.. Progress in Modern Biomedicine[J], 2008, 8(10): 1939—1941
[17]Maiorino M., Aumann K.D., Brigelius-Flohé R., et al.. Z. Ernahrungswiss.[J],1998, 37(Suppl 1): 118—121
[18]Maiorino M., Aumann K.D., Brigelius-Flohé R., et al.. Biol. Chem. Hoppe Seyler.[J], 1995, 376(11): 651—660
[19]Zheng K. Y., Board P. G., Fei X. F., et al.. Int. J. Biochem. Cell Biol.[J], 2008, 40(10): 2090—2097

Substrate-binding Sites of Human Selenium-containing Single Chain Abzyme by Quikchange Site-directed Mutagenesis

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