SPATIAL SPECIFICITY OF CHROMOPHORE ASSISTED LASER INACTIVATION OF PROTEIN FUNCTION
SPATIAL SPECIFICITY OF CHROMOPHORE ASSISTED LASER INACTIVATION OF PROTEIN FUNCTION
复制标题
DOI：
10.1016/s0006-3495(92)81902-1
复制
发表时间：
1992-04-01
影响因子：
3.4
通讯作者：
JAY, DG
中科院分区：
文献类型：
--
作者：
LINDEN, KG;LIAO, JC;JAY, DG
发色团辅助激光灭活(CALI)是一种选择性地灭活目标蛋白以阐明其在体内的功能的新技术。这种方法适用于一系列广泛的生物学问题。要明智地应用CALI，就需要对CALI机制的各个方面有所了解。CALI的一个关键问题是它的空间特异性，因为CALI可能会使邻近的非结合蛋白失活。我们在这里表明，CALI非常依赖于发色团和蛋白质之间的距离，因此在60埃以上没有明显的影响。使用抗体的CALI可以通过复合体使其他蛋白质失活，但对于每种中间蛋白质，其有效性大约降低四倍。这些数据表明，CALI是空间特异性的，不太可能对邻近的蛋白质造成损害。
Chromophore assisted laser inactivation (CALI) is a new technique that selectively inactivates proteins of interest to elucidate their in vivo functions. This method has application to a wide array of biological questions. An understanding of aspects of the mechanism of CALI is required for its judicious application. A critical concern for CALI is its spatial specificity because nonspecific inactivation of neighboring unbound proteins by CALI is a possibility. We show here that CALI is very dependent on the distance between the chromophore and the protein such that there is no significant effect beyond 60 angstrom. CALI using antibodies can inactivate other proteins through a complex but its efficacy decreases approximately fourfold for each intervening protein. These data imply that CALI is spatially specific and damage to neighboring proteins is unlikely.