8GHR

Structure of human ENPP1 in complex with variable heavy domain VH27.2

Summary for 8GHR

• Entry DOI: 10.2210/pdb8ghr/pdb
• EMDB information: 40047
• Descriptor: Ectonucleotide pyrophosphatase/phosphodiesterase family member 1, secreted form, Immunoglobulin gamma-1 heavy chain fusion, VH27, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (8 entities in total)
• Functional Keywords: phosphodiesterase, inhibitor, immune system
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 261723.97

Authors

Carozza, J.A.,Wang, H.,Solomon, P.E.,Wells, J.A.,Li, L. (deposition date: 2023-03-10, release date: 2023-08-02, Last modification date: 2025-05-21)

Primary citation

Solomon, P.E.,Bracken, C.J.,Carozza, J.A.,Wang, H.,Young, E.P.,Wellner, A.,Liu, C.C.,Sweet-Cordero, E.A.,Li, L.,Wells, J.A.
Discovery of VH domains that allosterically inhibit ENPP1.
Nat.Chem.Biol., 20:30-41, 2024

PubMed Abstract: Ectodomain phosphatase/phosphodiesterase-1 (ENPP1) is overexpressed on cancer cells and functions as an innate immune checkpoint by hydrolyzing extracellular cyclic guanosine monophosphate adenosine monophosphate (cGAMP). Biologic inhibitors have not yet been reported and could have substantial therapeutic advantages over current small molecules because they can be recombinantly engineered into multifunctional formats and immunotherapies. Here we used phage and yeast display coupled with in cellulo evolution to generate variable heavy (VH) single-domain antibodies against ENPP1 and discovered a VH domain that allosterically inhibited the hydrolysis of cGAMP and adenosine triphosphate (ATP). We solved a 3.2 Å-resolution cryo-electron microscopy structure for the VH inhibitor complexed with ENPP1 that confirmed its new allosteric binding pose. Finally, we engineered the VH domain into multispecific formats and immunotherapies, including a bispecific fusion with an anti-PD-L1 checkpoint inhibitor that showed potent cellular activity.

PubMed: 37400538
DOI: 10.1038/s41589-023-01368-5

Experimental method

ELECTRON MICROSCOPY (3.2 Å)