TY - JOUR
T1 - Inducing rigid local structure around the zinc-binding region by hydrophobic interactions enhances the homotrimerization and apoptotic activity of zinc-free TRAIL
AU - Lee, Hyun Wook
AU - Kim, Tae In
AU - Chan, Khon Huynh
AU - Kwon, Myung Hee
AU - Kim, Jeong Sun
AU - Jin, Moonsoo
AU - Kim, Yong Sung
N1 - Funding Information:
This work was supported by grants from Korea Research Foundation (205-2004-D00068 to Y.S.K.), the National R&D Program for Cancer Control, Ministry of Health & Welfare (0520110-1 to Y.S.K.), and the “GRRC” Project of Gyeonggi Provincial Government (to Y.S.K. and M.H.K.), Republic of Korea.
PY - 2007/10/26
Y1 - 2007/10/26
N2 - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), existing as homotrimer in solution, contains a unique zinc-binding site coordinated by three Cys230 residues at the tip of trimeric interface. TRAIL mutant with replacements of Cys230 with Ala (TRAILC230A) negligibly formed trimeric structure and showed no apoptotic activity. Here, to elucidate the relationship between the trimeric stability and the apoptotic activity of TRAILC230A, we rationally designed mutations to induce homotrimerization of TRAILC230A by substituting for the three residues involved in hydrogen bonding (Tyr183 and Tyr243) and putative repulsive electrostatic (Arg227) interactions at the buried trimeric interface into hydrophobic residues, like Y183F, Y243F, and R227I. The TRAILC230A-derived mutants exhibited enhanced homotrimerization, but only the mutants containing R227I exhibited significant apoptosis-inducing activity in cancer cells. These results, together with the induction of rigid local structure around the zinc-binding region by R227I in TRAILC230A, suggest that ordered, rigid structure around the zinc-binding region is critical for the homotrimerization and apoptotic activity of TRAIL.
AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), existing as homotrimer in solution, contains a unique zinc-binding site coordinated by three Cys230 residues at the tip of trimeric interface. TRAIL mutant with replacements of Cys230 with Ala (TRAILC230A) negligibly formed trimeric structure and showed no apoptotic activity. Here, to elucidate the relationship between the trimeric stability and the apoptotic activity of TRAILC230A, we rationally designed mutations to induce homotrimerization of TRAILC230A by substituting for the three residues involved in hydrogen bonding (Tyr183 and Tyr243) and putative repulsive electrostatic (Arg227) interactions at the buried trimeric interface into hydrophobic residues, like Y183F, Y243F, and R227I. The TRAILC230A-derived mutants exhibited enhanced homotrimerization, but only the mutants containing R227I exhibited significant apoptosis-inducing activity in cancer cells. These results, together with the induction of rigid local structure around the zinc-binding region by R227I in TRAILC230A, suggest that ordered, rigid structure around the zinc-binding region is critical for the homotrimerization and apoptotic activity of TRAIL.
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U2 - 10.1016/j.bbrc.2007.08.075
DO - 10.1016/j.bbrc.2007.08.075
M3 - Article
C2 - 17765202
AN - SCOPUS:34548392642
SN - 0006-291X
VL - 362
SP - 766
EP - 772
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -
