Biological and physicochemical evaluation of the conformational stability of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).
Tumor necrosis factor (TNF)-related, apoptosis-inducing ligand (Apo2L/TRAIL) has a unique homotrimeric structure, and its conformational stability is essential for its apoptotic activity. The conformational stability of a modified version of TRAIL(114-281) with two additional domains of histidine tag and isoleucine zipper [His-ILZ-TRAIL(114-281)] was evaluated in various pH environments according to three different biological or physicochemical considerations: cytotoxicity, antibody-binding affinity, and tertiary structure. The biological properties of His-ILZ-TRAIL(114-281) were the most stably maintained at pH 6.0. The physicochemical analyses (circular dichroism and fluorescence spectroscopy) demonstrate that its bioactivity loss by pH challenge was originated from its structural collapse as a homotrimer.
Youn
Yu Seok
YS
Drug Targeting Laboratory, College of Pharmacy, SungKyunKwan University, 300 Chonchon-dong, Jangan-ku, Suwon City, 440-746, Korea.
Shin
Min Jae
MJ
Chae
Su Young
SY
Jin
Cheng-Hao
CH
Kim
Tae Hyung
TH
Lee
Kang Choon
KC
D000915:Antibody Affinity / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D006207:Half-Life; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011487:Protein Conformation / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000737:chemistry* / Q000494:pharmacology; D013816:Thermodynamics; D013997:Time Factors
D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand