Article info Vol. 6  No. 4   pp.  92 ~ 95
Title Crystallization and preliminary X-ray analysis of API5-FGF2 complex
Authors Seoung Min Bong1 and Byung Il Lee1,2,*
Institutions 1Research institute, National Cancer Center, Goyang-si, Gyeonggi 10408, Republic of Korea, 2National Cancer Center Graduate School of Cancer Science and Policy, Goyang-si, Gyeonggi 10408, Republic *Correspondence: bilee@ncc.re.kr
Abstract API5 is a unique oncogenic, non-BIR type IAP nuclear protein and is up-regulated in several cancers. It exerts several functions, such as apoptosis inhibition, cell cycle progression, cancer immune escape, and anticancer drug resistance. Although structural studies of API have revealed that API5 mediates protein-protein interactions, its detailed molecular functions remain unknown. Since FGF2 is one of API5’s major interacting proteins, structural studies of the API5–FGF2 complex will provide insight into both proteins’ molecular function. We overexpressed and purified API5 and FGF2 in Escherichia coli and crystallized the API–FGF2 complex using polyethylene glycol (PEG) 6000 as a precipitant. Diffraction data were collected to a 2.7 Å resolution using synchrotron X-rays. Preliminary diffraction analysis revealed that the API5–FGF2 complex crystal belongs to the space group P212121 with the following unit cell parameters: a = 46.862, b = 76.523, c = 208.161 Å. One asymmetric unit with 49.9% solvent contains one API5-FGF2 complex. Molecular replacement calculation, using API5 and FGF2 coordinates, provided a clear electron density map for an API5–FGF2 complex.