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PDBsum entry 3hmx
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Cytokine/immune system
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PDB id
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3hmx
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Contents |
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295 a.a.
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176 a.a.
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214 a.a.
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217 a.a.
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* Residue conservation analysis
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PDB id:
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| Name: |
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Cytokine/immune system
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Title:
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Crystal structure of ustekinumab fab/il-12 complex
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Structure:
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Interleukin-12 subunit beta. Chain: a. Synonym: il-12b, il-12 subunit p40, cytotoxic lymphocyte maturation factor 40 kda subunit, clmf p40, nk cell stimulatory factor chain 2, nksf2. Engineered: yes. Interleukin-12 subunit alpha. Chain: b. Synonym: il-12a, il-12 subunit p35, cytotoxic lymphocyte maturation
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Source:
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Homo sapiens. Human. Organism_taxid: 9606. Gene: il12b, nksf2. Expressed in: homo sapiens. Expression_system_taxid: 9606. Expression_system_cell_line: hek 293. Gene: il12a, nksf1. Other_details: papain digested from mab.
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Resolution:
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3.00Å
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R-factor:
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0.215
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R-free:
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0.303
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Authors:
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J.Luo
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Key ref:
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J.Luo
et al.
(2010).
Structural basis for the dual recognition of IL-12 and IL-23 by ustekinumab.
J Mol Biol,
402,
797-812.
PubMed id:
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Date:
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29-May-09
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Release date:
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09-Jun-10
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PROCHECK
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Headers
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References
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P29460
(IL12B_HUMAN) -
Interleukin-12 subunit beta from Homo sapiens
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Seq:
Struc:
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328 a.a.
295 a.a.
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P29459
(IL12A_HUMAN) -
Interleukin-12 subunit alpha from Homo sapiens
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Seq:
Struc:
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219 a.a.
176 a.a.
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J Mol Biol
402:797-812
(2010)
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PubMed id:
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Structural basis for the dual recognition of IL-12 and IL-23 by ustekinumab.
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J.Luo,
S.J.Wu,
E.R.Lacy,
Y.Orlovsky,
A.Baker,
A.Teplyakov,
G.Obmolova,
G.A.Heavner,
H.T.Richter,
J.Benson.
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ABSTRACT
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Interleukin (IL)-12 and IL-23 are heterodimeric proinflammatory cytokines that
share a common p40 subunit, paired with p35 and p19 subunits, respectively. They
represent an attractive class of therapeutic targets for the treatment of
psoriasis and other immune-mediated diseases. Ustekinumab is a fully human
monoclonal antibody (mAb) that binds specifically to IL-12/IL-23p40 and
neutralizes human IL-12 and IL-23 bioactivity. The crystal structure of
ustekinumab Fab (antigen binding fragment of mAb), in complex with human IL-12,
has been determined by X-ray crystallography at 3.0 Å resolution. Ustekinumab
Fab binds the D1 domain of the p40 subunit in a 1:1 ratio in the crystal,
consistent with a 2 cytokines:1 mAb stoichiometry, as measured by isothermal
titration calorimetry. The structure indicates that ustekinumab binds to the
same epitope on p40 in both IL-12 and IL-23 with identical interactions.
Mutational analyses confirm that several residues identified in the
IL-12/IL-23p40 epitope provide important molecular binding interactions with
ustekinumab. The electrostatic complementarity between the mAb antigen binding
site and the p40 D1 domain epitope appears to play a key role in
antibody/antigen recognition specificity. Interestingly, this structure also
reveals significant structural differences in the p35 subunit and p35/p40
interface, compared with the published crystal structure of human IL-12,
suggesting unusual and potentially functionally relevant structural flexibility
of p35, as well as p40/p35 recognition. Collectively, these data describe unique
observations about IL-12p35 and ustekinumab interactions with p40 that account
for its dual binding and neutralization of IL-12 and IL-23.
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