skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Crystal structure of the m4-1BB/4-1BBL complex reveals an unusual dimeric ligand that undergoes structural changes upon 4-1BB receptor binding

Journal Article · · Journal of Biological Chemistry
ORCiD logo [1]; ORCiD logo [2];  [3];  [4]; ORCiD logo [5]
  1. La Jolla Inst. for Immunology (LJI), La Jolla, CA (United States)
  2. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  3. Kirin Kyowa Hakko Pharmaceutical Research, La Jolla, CA (United States)
  4. La Jolla Inst. for Immunology (LJI), La Jolla, CA (United States); Univ. of California San Diego, La Jolla, CA (United States)
  5. La Jolla Inst. for Immunology (LJI), La Jolla, CA (United States); Ghent Univ., Ghent (Belgium)
+ Show Author Affiliations

The interaction between the receptor 4-1BB and its ligand 4-1BBL provides co-stimulatory signals for T-cell activation and proliferation. However, differences in the mouse and human molecules might result in differential engagement of this pathway. Here, we report the crystal structure of mouse 4-1BBL and of the mouse 4-1BB/4-1BBL complex, which together provided insights into the molecular mechanism by which m4-1BBL and its cognate receptor recognize each other. Unlike all human or mouse tumor necrosis factor ligands that form noncovalent and mostly trimeric assemblies, the m4-1BBL structure formed a disulfide-linked dimeric assembly. The structure disclosed that certain differences in the amino acid composition along the intramolecular interface, together with two specific residues (Cys-246 and Ser-256) present exclusively in m4-1BBL, are responsible for this unique dimerization. Unexpectedly, upon m4-1BB binding, m4-1BBL undergoes structural changes within each protomer; moreover, the individual m4-1BBL protomers rotate relative to each other, yielding a dimerization interface with more inter-subunit interactions. We also observed that in the m4-1BB/4-1BBL complex, each receptor monomer binds exclusively to a single ligand subunit with contributions of cysteine-rich domain 1 (CRD1), CRD2, and CRD3. Furthermore, structure-guided mutagenesis of the binding interface revealed that novel binding interactions with the GH loop, rather than the DE loop, are energetically critical and define the m4-1BB receptor selectivity for m4-1BBL. As a result, a comparison with the human 4-1BB/4-1BBL complex highlighted several differences between the ligand- and receptor-binding interfaces, providing an explanation for the absence of inter-species cross-reactivity between human and mouse 4-1BB and 4-1BBL molecules.

Research Organization:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Organization:
USDOE
Grant/Contract Number:
AC02-76SF00515; AI110929; P41GM103393
OSTI ID:
1506962
Journal Information:
Journal of Biological Chemistry, Vol. 294, Issue 6; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 11 works
Citation information provided by
Web of Science

References (34)

A casein kinase I motif present in the cytoplasmic domain of members of the tumour necrosis factor ligand family is implicated in `reverse signalling' journal April 1999
Monoclonal antibodies against the 4-1BB T-cell activation molecule eradicate established tumors journal June 1997
Overview of the CCP 4 suite and current developments journal March 2011
The Crystal Structure of the Costimulatory OX40-OX40L Complex journal August 2006
Structural basis for ligand-mediated mouse GITR activation journal January 2008
Molecular replacement – historical background journal September 2001
Triggering Cell Death journal October 1999
Crystal structure of the human 4-1BB/4-1BBL complex journal May 2018
Crystal Structure of TRAIL-DR5 Complex Identifies a Critical Role of the Unique Frame Insertion in Conferring Recognition Specificity journal July 2000
Galectin-9 controls the therapeutic activity of 4-1BB–targeting antibodies journal June 2014
The TNF and TNF Receptor Superfamilies journal February 2001
Crystallographic and Mutational Analysis of the CD40-CD154 Complex and Its Implications for Receptor Activation journal February 2011
A graphical user interface to the CCP 4 program suite journal June 2003
Evolution of GITRL immune function: Murine GITRL exhibits unique structural and biochemical properties within the TNF superfamily journal January 2008
Human and Mouse CD137 Have Predominantly Different Binding CRDs to Their Respective Ligands journal January 2014
Species Difference of CD137 Ligand Signaling in Human and Murine Monocytes journal January 2011
The molecular architecture of the TNF superfamily journal January 2002
The Structure of the Trimer of Human 4-1BB Ligand Is Unique among Members of the Tumor Necrosis Factor Superfamily journal December 2009
Crystal structure of murine 4-1BB and its interaction with 4-1BBL support a role for galectin-9 in 4-1BB signaling journal December 2017
CD137 ligand reverse signaling has multiple functions in human dendritic cells during an adaptive immune response journal April 2008
T-cell intrinsic effects of GITR and 4-1BB during viral infection and cancer immunotherapy: Effects of GITR and 4-1BB on T cells journal October 2011
4-1BB T-cell antigen binds to mature B cells and macrophages, and costimulates anti-μ-primed splenic B cells journal February 1994
Crystal Structure of Human RANKL Complexed with Its Decoy Receptor Osteoprotegerin journal June 2012
Sequence, structure, function, immunity: structural genomics of costimulation journal May 2009
MOLREP an Automated Program for Molecular Replacement journal December 1997
4-1BB Ligand Signaling to T Cells Limits T Cell Activation journal November 2014
Role of 4-1BB in immune responses journal December 1998
Crystal structures of the human 4-1BB receptor bound to its ligand 4-1BBL reveal covalent receptor dimerization as a potential signaling amplifier journal May 2018
Modularity in the TNF-receptor family journal February 1998
4-1BB and Ox40 Are Members of a Tumor Necrosis Factor (TNF)-Nerve Growth Factor Receptor Subfamily That Bind TNF Receptor-Associated Factors and Activate Nuclear Factor κB journal January 1998
Assembly and structural properties of glucocorticoid-induced TNF receptor ligand: Implications for function journal November 2007
Deciphering CD137 (4-1BB) signaling in T-cell costimulation for translation into successful cancer immunotherapy journal February 2016
PARP1 exhibits enhanced association and catalytic efficiency with γH2A.X-nucleosome journal December 2019
Overview of the CCP4 suite and current developments. text January 2011

Cited By (4)

Tumor-targeted 4-1BB agonists for combination with T cell bispecific antibodies as off-the-shelf therapy journal June 2019
A universal reporter cell line for bioactivity evaluation of engineered cytokine products journal February 2020
A universal reporter cell line for bioactivity evaluation of engineered cytokine products text January 2020
A Systematic Test of Receptor Binding Kinetics for Ligands in Tumor Necrosis Factor Superfamily by Computational Simulations journal March 2020

Similar Records

Crystal structures of the human 4-1BB receptor bound to its ligand 4-1BBL reveal covalent receptor dimerization as a potential signaling amplifier
Journal Article · Wed May 02 00:00:00 EDT 2018 · Journal of Biological Chemistry · OSTI ID:1506962
+1 more
Structure of the 4-1BB/4-1BBL complex and distinct binding and functional properties of utomilumab and urelumab
Journal Article · Thu Nov 08 00:00:00 EST 2018 · Nature Communications · OSTI ID:1506962
+13 more
Genomic organization and chromosomal localization of the T-cell antigen 4-1BB
Journal Article · Tue Mar 01 00:00:00 EST 1994 · Journal of Immunology; (United States) · OSTI ID:1506962
+1 more

Related Subjects

60 APPLIED LIFE SCIENCES
tumor necrosis factor (TNF)
protein–protein interaction
protein structure
cell surface receptor
X-ray crystallography
4-1BB
4-1BB ligand
CD137
CD137L
TNFRSF9
immune cell