2ZPX

TNF Receptor Subtype One-selective TNF Mutant with Antagonistic Activity; R1antTNF-T8

Summary for 2ZPX

• Entry DOI: 10.2210/pdb2zpx/pdb
• Related: 1TNF 1TNR 2EA7 2ZJC
• Descriptor: Tumor necrosis factor (2 entities in total)
• Functional Keywords: tumor necrosis factor, trimer, antagonistic activity, tnfr1 specific, phage display system, cytokine, cell membrane, lipoprotein, membrane, myristate, phosphoprotein, polymorphism, secreted, signal-anchor, transmembrane
• Biological source: Homo sapiens (Human)
• Cellular location: Cell membrane; Single-pass type II membrane protein. Tumor necrosis factor, soluble form: Secreted: P01375
• Total number of polymer chains: 3
• Total formula weight: 51850.31

Authors

Mukai, Y.,Nakamura, T.,Yamagata, Y.,Tsutsumi, Y. (deposition date: 2008-07-29, release date: 2009-03-24, Last modification date: 2024-10-16)

Primary citation

Mukai, Y.,Nakamura, T.,Yoshioka, Y.,Shibata, H.,Abe, Y.,Nomura, T.,Taniai, M.,Ohta, T.,Nakagawa, S.,Tsunoda, S.,Kamada, H.,Yamagata, Y.,Tsutsumi, Y.
Fast binding kinetics and conserved 3D structure underlie the antagonistic activity of mutant TNF: useful information for designing artificial proteo-antagonists
J.Biochem., 146:167-172, 2009

PubMed Abstract: Tumour necrosis factor (TNF) is an important cytokine that induces an inflammatory response predominantly through the TNF receptor-1 (TNFR1). A crucial strategy for the treatment of many autoimmune diseases, therefore, is to block the binding of TNF to TNFR1. We previously identified a TNFR1-selective antagonistic mutant TNF (R1antTNF) from a phage library containing six randomized amino acid residues at the receptor-binding site (amino acids 84-89). Two R1antTNFs, R1antTNF-T2 (A84S, V85T, S86T, Y87H, Q88N and T89Q) and R1antTNF-T8 (A84T, V85P, S86A, Y87I, Q88N and T89R), were successfully isolated from this library. Here, we analysed R1antTNF-T8 using surface plasmon resonance spectroscopy and X-ray crystallography to determine the mechanism underlying the antagonistic activity of R1antTNF. The kinetic association/dissociation parameters of R1antTNF-T8 were higher than those of wild-type TNF, indicating more rapid bond dissociation. X-ray crystallographic analysis suggested that the binding mode of the T89R mutation changed from a hydrophobic to an electrostatic interaction, which may be responsible for the antagonistic behaviour of R1antTNF. Knowledge of these structure-function relationships will facilitate the design of novel TNF inhibitors based on the cytokine structure.

PubMed: 19386778
DOI: 10.1093/jb/mvp065

Experimental method

X-RAY DIFFRACTION (2.83 Å)