Figure 5

Association of PIG-2 and YWHAH. (A) Mapping of PIG-2 binding domain of YWHAH. Five GST fusion constructs, Full-YWHAHp1-247, YWHAHp1-100, YWHAHp1–80, YWHAHp1–60 and YWHAHp81–247 were prepared using YWHAH cDNA as a template DNA. The GST pull-down assay showed that Three GST-YWHAH constructs, Full-YWHAHp1–247, YWHAHp1–100 and YWHAHp1–80 bind YWHAH, but not YWHAHp1–60 and YWHAHp60–247. The YWHAH binding site for PIG-2 was delineated to be residues 61–80 (black box). (B) Mapping of YWHAH binding domain of PIG-2. Four GST fusion constructs, Full-PIG-2p1-184, PIG-2p1-144, PIG-2p1-100 and PIG-2p1-67 were prepared using PIG-2 cDNA as a template DNA. The GST pull-down assay showed all constructs, The PIG-2 binding site for YWHAH was delineated to be residues 1–67. Gray box is DAN domain. (C) Schematic diagram shows X-ray crystallography of YWHAH protein. PIG-2 protein binding site was indicated by arrow.