Crystal Structures of the Pro-Inflammatory Cytokine Interleukin-23 and Its Complex with a High-Affinity Neutralizing Antibody

Beyer, Brian M.; Ingram, Richard; Ramanathan, Lata; Reichert, Paul; Le, Hung V.; Madison, Vincent; Orth, Peter

doi:10.1016/j.jmb.2008.08.001

Cited by 51 publications

(54 citation statements)

References 32 publications

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“…14,21,25,26 Antibodies produced in this format are easily purified and tested using functional in vitro and cellbased assays. For example, the cell-free produced anti-hIL-23 scFv bound specifically to the p19 subunit of human IL-23, 27 with a K D of 10 nM, as measured by surface plasmon resonance (SPR; Fig. 2D).…”

Section: Resultsmentioning

confidence: 96%

“…A strong T7 transcription terminator hairpin prevents elongated transcripts and 3'-terminal exonucleolytic degradation of the mRNA. 43 Genes coding for an scFv that binds the p19 subunit of human IL-23, 27 a Fab that binds to human IL13R-1α 25 with a C-terminal hinge containing c-myc-tag and His-tags, and trastuzumab 44 IgG1 HC and LC were codon optimized and synthesized (DNA 2.0, Menlo Park, CA).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system

Yin¹,

Garces²,

Yang³

et al. 2012

mAbs

129

157

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Section: Resultsmentioning

confidence: 96%

Section: Methodsmentioning

confidence: 99%

Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system

Yin¹,

Garces²,

Yang³

et al. 2012

mAbs

129

157

View full text Add to dashboard Cite

show abstract

“…Although it has been suggested that both IL-17 and IL-23 may be important mediators in chronic Lyme disease, this was not confirmed in functional studies (5). In rheumatoid arthritis (RA), it was already demonstrated that the IL-17/IL-23 axis plays an important role and that individuals with the single nucleotide polymorphism in IL-23R are protected against the development of RA (16).…”

Section: Discussionmentioning

confidence: 99%

Role of Interleukin-23 (IL-23) Receptor Signaling for IL-17 Responses in Human Lyme Disease

et al. 2011

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“…Each has its advantages and disadvantages. X-ray diffraction provides direct structural information that can define the binding interface at atomic resolution (12,13) but this requires the difficult crystallization of the unperturbed complex. NMR spectroscopy offers a similar level of structural detail but is limited to relatively small proteins that can be expressed with 15 Nand 13 C-labeled residues and prepared in large amounts at high concentration.…”

mentioning

confidence: 99%

High-resolution epitope mapping by HX MS reveals the pathogenic mechanism and a possible therapy for autoimmune TTP syndrome

Casina

Mao

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Acquired thrombotic thrombocytopenic purpura (TTP), a thrombotic disorder that is fatal in almost all cases if not treated promptly, is primarily caused by IgG-type autoantibodies that inhibit the ability of the ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) metalloprotease to cleave von Willebrand factor (VWF). Because the mechanism of autoantibody-mediated inhibition of ADAMTS13 activity is not known, the only effective therapy so far is repeated whole-body plasma exchange. We used hydrogen-deuterium exchange mass spectrometry (HX MS) to determine the ADAMTS13 binding epitope for three representative human monoclonal autoantibodies, isolated from TTP patients by phage display as tethered single-chain fragments of the variable regions (scFvs). All three scFvs bind the same conformationally discontinuous epitopic region on five small solvent-exposed loops in the spacer domain of ADAMTS13. The same epitopic region is also bound by most polyclonal IgG autoantibodies in 23 TTP patients that we tested. The ability of ADAMTS13 to proteolyze VWF is impaired by the binding of autoantibodies at the epitopic loops in the spacer domain, by the deletion of individual epitopic loops, and by some local mutations. Structural considerations and HX MS results rule out any disruptive structure change effect in the distant ADAMTS13 metalloprotease domain. Instead, it appears that the same ADAMTS13 loop segments that bind the autoantibodies are also responsible for correct binding to the VWF substrate. If so, the autoantibodies must prevent VWF proteolysis simply by physically blocking normal ADAMTS13 to VWF interaction. These results point to the mechanism for autoantibody action and an avenue for therapeutic intervention.ADAMTS13 | von Willebrand factor | thrombotic thrombocytopenic purpura | hydrogen exchange | autoimmunity A cquired thrombotic thrombocytopenic purpura (TTP) is characterized by severe thrombocytopenia and microangiopathic hemolytic anemia, resulting from disseminated microvascular thrombosis. Patients with TTP may suffer from widespread organ damage, resulting in death if not aggressively treated (1, 2). In most patients, thrombotic microangiopathy is caused by IgG-type autoantibodies against the plasma metalloprotease ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) (3-5). ADAMTS13 regulates the function of the multidomain von Willebrand factor (VWF) (∼600 to ∼20,000 kDa) by cleaving it at the central A2 domain to regulate platelet-induced blood clot formation (3, 4).Immunological studies show that many acquired TTP patients with low plasma ADAMTS13 activity (less than 10%) harbor IgG autoantibodies that bind ADAMTS13 especially at the Cysrich and/or the spacer domain (4, 6). Deletion of these domains or grouped mutations therein can eliminate the binding of polyclonal anti-ADAMTS13 IgGs derived from many patients (4, 7-11). However, the exact ADAMTS13 binding sites of these autoantibodies are not known. Current treatment...

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Crystal Structures of the Pro-Inflammatory Cytokine Interleukin-23 and Its Complex with a High-Affinity Neutralizing Antibody

Cited by 51 publications

References 32 publications

Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system

Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system

Role of Interleukin-23 (IL-23) Receptor Signaling for IL-17 Responses in Human Lyme Disease

High-resolution epitope mapping by HX MS reveals the pathogenic mechanism and a possible therapy for autoimmune TTP syndrome

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