[1]曲小龙,胡厚源,李敏,等.抗炎、抗凝双效融合蛋白TAP-SSL5表达载体的构建及其功能研究[J].第三军医大学学报,2010,32(01):5-8.
 Qu Xiaolong,Hu Houyuan,Li Min,et al.Construction of a novel fusion protein TAP-SSL5 and evaluation of its function as anti-inflammatory and anticoagulant protein in vitro[J].J Third Mil Med Univ,2010,32(01):5-8.
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抗炎、抗凝双效融合蛋白TAP-SSL5表达载体的构建及其功能研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第01期
页码:
5-8
栏目:
论著
出版日期:
2010-01-15

文章信息/Info

Title:
Construction of a novel fusion protein TAP-SSL5 and evaluation of its function as anti-inflammatory and anticoagulant protein in vitro
作者:
曲小龙胡厚源李敏黄德兴梁华
第三军医大学:西南医院心血管内科,重庆市介入心脏病学研究所,护理系心理学教研室;BakerIDI Heart & Diabetes Institute
Author(s):
Qu Xiaolong Hu Houyuan Li Min Huang Dexing3 Liang Hua
Department of Cardiology, Southwest Hospital, Department of Psychology, College of Nursing, Third Military Medical University, Chongqing, 400038, China; BakerIDI Heart & Diabetes Institute, Melbourne 8008, Australia
关键词:
金黄色葡萄球菌超抗原样蛋白-5蜱抗凝血肽融合蛋白P-选择素糖蛋白配体-1凝血因子Xa
Keywords:
staphylococcal superantigen-like protein-5 tick anticoagulant peptide fusion protein P-selectin glycoprotein ligand-1 factor Xa
分类号:
R394-33;R345.13;R972.6
文献标志码:
A
摘要:
目的  构建一种具有抗炎和抗凝双效功能的融合蛋白,并对其功能进行初步鉴定。 方法   将重组金黄色葡萄球菌超抗原样蛋白-5 (staphylococcal superantigenlike protein-5, SSL5)与蜱抗凝血肽(tick anticoagulant peptide, TAP)融合,从金黄色葡萄球NCTC8325菌株中克隆SSL5基因; 经DNA重组技术,将编码pelB前导序列、SSL5和TAP的基因重组并克隆于原核表达载体pHOG21中,再转染于大肠杆菌BL21中扩增表达,制备融合蛋白SSL5-TAP或TAP-SSL5,采用流式细胞仪检测融合蛋白是否和SSL5一样,具有与粒细胞表面P-选择素糖蛋白配体-1(P-selectin glycoprotein ligand-1, PSGL-1)结合的特性;采用反应底物法在体外检测融合蛋白对活化的凝血因子X(factor Xa,FXa)活性的抑制作用。  结果  TAP-SSL5保留了SSL5与PSGL-1结合的特性,并且融合蛋白TAP-SSL5可显著抑制FXa的活性(P<0.01),抑制率达39.5%。 结论 融合蛋白TAP-SSL5是一种以PSGL-1为靶向的新型抗炎、抗凝双效分子。
Abstract:
Objective To construct a double-effect fusion protein functioning as an anti-inflammatory and anticoagulant, which can inhibit the inflammatory response and thrombosis simultaneously in atherosclerosis lesions. Methods Firstly, staphylococcal superantigen-like protein-5 (SSL5) gene sequence was cloned by PCR from Staphylococcus aureus (strain NCTC8325). Secondly, N-terminal pelB leading sequence, SSL5 gene sequence, tick anticoagulant peptide (TAP) gene sequence, with C-terminal both c-Myc and His6 tags were cloned into the pHOG21 expression vector using recombined DNA technique, and the obtained recombinant were further transformed into E. coli BL21(DE3) cells to express the fusion protein TAP-SSL5. Flow cytometry was used to investigate whether the fusion protein could bind with P-selectin glycoprotein ligand-1 (PSGL-1) on monocytes as SSL5. Finally, the inhibitory effect of TAP-SSL5 on factor Xa (FXa) activity were detected by chromogenic substrate assay in vitroResults TAP-SSL5 fusion protein binded to PSGL-1 on HL60 monocytes, and inhibited the FXa activity in vitro as well, with an inhibitory rate of 39.5% (P<0.01).  Conclusion  The fusion protein TAP-SSL5 is a novel molecule targeting PSGL-1, which has an double-effects of anti-inflammation and anticoagulation.

参考文献/References:

曲小龙, 胡厚源, 李敏, 等.抗炎、抗凝双效融合蛋白TAP-SSL5表达载体的构建及其功能研究[J]. 第三军医大学学报,2010,32(1):5-8.

相似文献/References:

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 Liang Hua,Qu Xiaolong,Hu Houyuan,et al.Staphylococcal superantigen-like protein-5 inhibits adhesion of human umbilical cord blood-derived endothelial progenitor cells to P-selectin-coated surface[J].J Third Mil Med Univ,2011,33(01):545.
[2]程彦,房兆飞,曲小龙,等.融合蛋白TAP-SSL5对人血小板功能的影响[J].第三军医大学学报,2012,34(06):477.
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[4]龚丽莎,房兆飞,胡厚源,等.融合蛋白TAP-SSL5对血小板与粒细胞结合的影响[J].第三军医大学学报,2013,35(08):754.
 Gong Lisha,Fang Zhaofei,Hu Houyuan,et al.Effect of TAP-SSL5 fusion protein on binding of human platelets to neutrophils[J].J Third Mil Med Univ,2013,35(01):754.
[5]刘成海,彭松,胡厚源,等.融合蛋白TAP-SSL5对血小板微粒与THP-1细胞结合及Mac-1活化的影响[J].第三军医大学学报,2014,36(09):864.
 Liu Chenghai,Peng Song,Hu Houyuan,et al.Fusion protein TAP-SSL5 suppresses binding of platelet microparticles to THP-1 cells and activation of Mac-1[J].J Third Mil Med Univ,2014,36(01):864.

更新日期/Last Update: 2010-01-08