利用酵母杂交系统克隆一种新鉴定出的心脏特异性肌钙蛋白I同工型，该同工型缺乏肌钙蛋白T结合部分。
Cloning of a newly identified heart-specific troponin I isoform, which lacks the troponin T binding portion, using the yeast hybrid system.
作者信息
Suzuki Hideaki, Arakawa Yasuhiro, Ito Masaki, Yamada Hisashi, Horiguchi-Yamada Junko
机构信息
Departments of Internal Medicine and.
出版信息
Exp Clin Cardiol. 2006 Spring;11(1):4-7.
OBJECTIVE
To elucidate the molecular pathogenesis behind increased levels of laminin in cardiac muscle cells in cardiomyopathy by using a yeast hybrid screen. The present study reports the cloning of a newly identified heart-specific troponin I isoform, which is putatively linked to laminin. Future studies will explore the functional significance of this connection.
METHODS
Yeast two-hybrid screen analysis was performed using MLF1-interacting protein (amino acids 1 to 318) as bait. The human heart complementary DNA library was screened by using the yeast-mating method for overnight culture.
RESULTS
Two final positive clones from the heart library were isolated. These two clones encoded the same protein, a short isoform of human cardiac troponin I (TnI) that lacked TnI exons 5 and 6. The TnI isoform has a heart-specific expression pattern and it shares several sequence features with human cardiac TnI; however, it lacks the troponin T binding portion.
CONCLUSION
The heart-specific segment of the human cardiac TnI isoform shares several sequence features with human cardiac TnI, but it lacks the troponin T binding portion. These results suggest that the heart-specific TnI isoform may be involved in cardiac development and disease.
目的
通过酵母杂交筛选阐明心肌病中心肌细胞层粘连蛋白水平升高背后的分子发病机制。本研究报告了一种新鉴定的心脏特异性肌钙蛋白I同工型的克隆，其可能与层粘连蛋白有关。未来的研究将探索这种联系的功能意义。
方法
以MLF1相互作用蛋白（氨基酸1至318）为诱饵进行酵母双杂交筛选分析。用人心脏互补DNA文库通过酵母交配法进行过夜培养筛选。
结果
从心脏文库中分离出两个最终的阳性克隆。这两个克隆编码相同的蛋白质，即人心脏肌钙蛋白I（TnI）的一种短同工型，其缺少TnI外显子5和6。该TnI同工型具有心脏特异性表达模式，并且与人心肌TnI具有若干序列特征；然而，它缺少肌钙蛋白T结合部分。
结论
人心脏TnI同工型的心脏特异性片段与人心肌TnI具有若干序列特征，但缺少肌钙蛋白T结合部分。这些结果表明，心脏特异性TnI同工型可能参与心脏发育和疾病。
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