Cysteine 230 is essential for the structure and activity of the cytotoxic ligand TRAIL.
Unlike other tumor necrosis factor family members, the cytotoxic ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/Apo-2L contains an unpaired cysteine residue (Cys(230)) in its receptor-binding domain. Here we show that the biological activity of both soluble recombinant TRAIL and cell-associated, full-length TRAIL is critically dependent on the presence of Cys(230). Mutation of Cys(230) to alanine or serine strongly affected its ability to kill target cells. Binding to its receptors was decreased by at least 200-fold, and the stability of its trimeric structure was reduced. In recombinant TRAIL, Cys(230) was found engaged either in interchain disulfide bridge formation, resulting in poorly active TRAIL, or in the chelation of one zinc atom per TRAIL trimer in the active, pro-apoptotic form of TRAIL.
Bodmer
J L
JL
Institute of Biochemistry, University of Lausanne and Laboratoire Cantonal, CH-1066 Epalinges, Switzerland.
Meier
P
P
Tschopp
J
J
Schneider
P
P
D000595:Amino Acid Sequence; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D053221:TNF-Related Apoptosis-Inducing Ligand; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D015032:Zinc / Q000737:chemistry
D051017:Apoptosis Regulatory Proteins; D004220:Disulfides; D008562:Membrane Glycoproteins; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; D015032:Zinc; D003545:Cysteine