2HH0

Structure of an Anti-PrP Fab, P-Clone, in Complex with its Cognate Bovine Peptide Epitope.

2HH0 の概要

• エントリーDOI: 10.2210/pdb2hh0/pdb
• 分子名称: Light Chain, P-Clone Fab, Chimera, Heavy Chain, P-Clone Fab, Chimera, Prion protein, ... (4 entities in total)
• 機能のキーワード: prion, prp, fab, immune system
• 由来する生物種: Mus musculus, Homo sapiens (house mouse, human); 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 47366.76

構造登録者

Kanyo, Z.K. (登録日: 2006-06-27, 公開日: 2006-12-26, 最終更新日: 2024-11-20)

主引用文献

Luginbuhl, B.,Kanyo, Z.,Jones, R.M.,Fletterick, R.J.,Prusiner, S.B.,Cohen, F.E.,Williamson, R.A.,Burton, D.R.,Pluckthun, A.
Directed evolution of an anti-prion protein scFv fragment to an affinity of 1 pM and its structural interpretation
J.Mol.Biol., 363:75-97, 2006

PubMed Abstract: Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative prion disease affecting cattle that is transmissible to humans, manifesting as a variant of Creutzfeldt-Jakob disease (vCJD) likely following the consumption of meat contaminated with BSE prions. High-affinity antibodies are a prerequisite for the development of simple, highly sensitive and non-invasive diagnostic tests that are able to detect even small amounts of the disease-associated PrP conformer (PrP(Sc)). We describe here the affinity maturation of a single-chain Fv antibody fragment with a binding affinity of 1 pM to a peptide derived from the unstructured region of bovine PrP (BoPrP (90-105)). This is the tightest peptide-binding antibody reported to date and may find useful application in diagnostics, especially when PrP(Sc) is pretreated by denaturation and/or proteolysis for peptide-like presentation. Several rounds of directed evolution and off-rate selection with ribosome display were performed using an antibody library generated from a single PrP binder with error-prone PCR and DNA-shuffling. As the correct determinations of affinities in this range are not straightforward, competition biosensor techniques and KinExA methods were both applied and compared. Structural interpretation of the affinity improvement was performed based on the crystal structure of the original prion binder in complex with the BoPrP (95-104) peptide by modeling the corresponding mutations.

PubMed: 16962610
DOI: 10.1016/j.jmb.2006.07.027

実験手法

X-RAY DIFFRACTION (2.85 Å)