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| Prokaryotic Expression, Purification, Antibody Production of a NH2 -Terminal Fragment of mCLCA3 Protein and Analysis of mClCA3 Protein Expression in Asthmatic Mouse Lung | |
| 作者姓名： | HOU Xia WU Qing-tian ZHANG Yu-ping ZHU Na LIU Yan-li FENG Xue-chao MA Tong-hui |
| 作者单位： | HOU Xia1，2，WU Qing-tian1，2，ZHANG Yu-ping2，ZHU Na1，LIU Yan-li1，FENG Xue-chao1 and MA Tong-hui11.Membrane Channel Research Laboratory，Northeast Normal University，Changchun 130024，P.R.China;2.School of Basic Medicine，Jiamusi University，Jiamusi 154002，P.R.China |
| 基金项目： | Supported by the Distinguished Young Scholars Fund of China(No.30325011)，the National Natural Science Foundation of China(Nos.30470405and30670477)，Distinguished Young Scholars Fund of Jilin Province,China(No.20030112)，Excellent Young Teachers Program of MOE,China. |
| 摘 要： | mCLCA3 is a member of calcium activated chloride channel(CACC)family that may play an important role in mucin packaging and secretion in asthmatic and cystic fibrosis lung.To study the protein structure and expression of mCLCA3 in asthmatic mouse lung,an N-terminal 269 amino acid peptide of mCLCA3 was expressed in E.coli,purified to homogeneity and rabbit polyclonal antibodies against this peptide were generated.Immunohistochemistry of asthmatic mouse lung using the antibody indicated exclusive mCLCA3 expression in mucin granules of goblet cells in airway surface and lumen.Immunoblot analysis of lavage fluid from asthmatic mouse lung revealed a single 90 kDa protein form of mClCA3.The results demonstrate that the 90 kDa N-terminal peptide,neither the full-length protein nor the reported N-terminal 35 kDa cleaved form of mClCA3 is the major functional form involved in the packaging and exocytosis of mucin granules in asthmatic goblet cells. |
| 关 键 词： | 哮喘 肺 mCLCA3蛋白 原核表达 抗体 |
| 文章编号： | 1005-9040（2007）-06-688-05 |
| 收稿时间： | 2007-05-16 |
Prokaryotic Expression, Purification, Antibody Production of a NH2-Terminal Fragment of mCLCA3 Protein and Analysis of mClCA3 Protein Expression in Asthmatic Mouse Lung |
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| HOU Xia WU Qing-tian ZHANG Yu-ping ZHU Na LIU Yan-li FENG Xue-chao MA Tong-hui.Prokaryotic Expression, Purification, Antibody Production of a NH2 -Terminal Fragment of mCLCA3 Protein and Analysis of mClCA3 Protein Expression in Asthmatic Mouse Lung[J].Chemical Research in Chinese University,2007,23(6):688-692. | |
| Authors: | Xia HOU Qing-tian WU Yu-ping ZHANG Na ZHU Yan-li LIU Xue-chao FENG Tong-hui MA |
| Institution: | aMembrane Channel Research Laboratory, Northeast Normal University, Changchun 130024, P. R. China bSchool of Basic Medicine, Jiamusi University, Jiamusi 154002, P. R. China |
| Abstract: | mCLCA3 is a member of calcium activated chloride channel(CACC)family that may play an important role in mucin packaging and secretion in asthmatic and cystic fibrosis lung.To study the protein structure and expression of mCLCA3 in asthmatic mouse lung,an N-terminal 269 amino acid peptide of mCLCA3 was expressed in E.coli,purified to homogeneity and rabbit polyclonal antibodies against this peptide were generated.Immunohistochemistry of asthmatic mouse lung using the antibody indicated exclusive mCLCA3 expression in mucin granules of goblet cells in airway surface and lumen.Immunoblot analysis of lavage fluid from asthmatic mouse lung revealed a single 90 kDa protein form of mClCA3.The results demonstrate that the 90 kDa N-terminal peptide,neither the full-length protein nor the reported N-terminal 35 kDa cleaved form of mClCA3 is the major functional form involved in the packaging and exocytosis of mucin granules in asthmatic goblet cells. |
| Keywords: | mCLCA3 Prokaryotic expression Purification Antibody production Asthma |
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