Author: Wenbin Ji; Yibo Luo; Ejaz Ahmad; Song-Tao Liu
Title: Coordination between discrete Mitotic Arrest Deficient 1 (MAD1) domains is required for efficient mitotic checkpoint signaling Document date: 2017_11_1
ID: i4yquw4k_10
Snippet: We attempted to further define the regions on MAD1-NTD or CTD responsible for association with MAD2. Several MAD1-NTD or CTD truncations produced either insoluble or heavily degraded proteins (data not shown). However, testing with MAD1-NTD truncations that we were able to purify, including MAD1 , MAD1 , and MAD1 (327-488) , revealed dramatically reduced MAD2 binding capability of these fragments (Fig. 2d ). The globular subdomain (638-718 residu.....
Document: We attempted to further define the regions on MAD1-NTD or CTD responsible for association with MAD2. Several MAD1-NTD or CTD truncations produced either insoluble or heavily degraded proteins (data not shown). However, testing with MAD1-NTD truncations that we were able to purify, including MAD1 , MAD1 , and MAD1 (327-488) , revealed dramatically reduced MAD2 binding capability of these fragments (Fig. 2d ). The globular subdomain (638-718 residues) of MAD1-CTD retained approximately half MAD2 binding capacity of MAD1-CTD while the coiled coil subdomain in the CTD showed only residual binding (Fig. 2e) . These combined results suggested that the integrity of MAD1-NTD or CTD is crucial for binding to O-MAD2 or C-MAD2.
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