7KZ7

Crystals Structure of the Mutated Protease Domain of Botulinum Neurotoxin X (X4130B1).

7KZ7 の概要

• エントリーDOI: 10.2210/pdb7kz7/pdb
• 分子名称: Botulinum neurotoxin type X, ZINC ION, 1,2-ETHANEDIOL, ... (5 entities in total)
• 機能のキーワード: structural genomics, center for structural genomics of infectious diseases, csgid, neurotoxin x, protease domain, toxin
• 由来する生物種: Clostridium botulinum
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 51464.28

構造登録者

Blum, T.R.,Liu, H.,Packer, M.S.,Xiong, X.,Lee, P.G.,Zhang, S.,Richter, M.,Minasov, G.,Satchell, K.J.F.,Dong, M.,Liu, D.R.,Center for Structural Genomics of Infectious Diseases (CSGID) (登録日: 2020-12-10, 公開日: 2020-12-23, 最終更新日: 2023-10-18)

主引用文献

Blum, T.R.,Liu, H.,Packer, M.S.,Xiong, X.,Lee, P.G.,Zhang, S.,Richter, M.,Minasov, G.,Satchell, K.J.F.,Dong, M.,Liu, D.R.
Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity.
Science, 371:803-810, 2021

PubMed Abstract: Although bespoke, sequence-specific proteases have the potential to advance biotechnology and medicine, generation of proteases with tailor-made cleavage specificities remains a major challenge. We developed a phage-assisted protease evolution system with simultaneous positive and negative selection and applied it to three botulinum neurotoxin (BoNT) light-chain proteases. We evolved BoNT/X protease into separate variants that preferentially cleave vesicle-associated membrane protein 4 (VAMP4) and Ykt6, evolved BoNT/F protease to selectively cleave the non-native substrate VAMP7, and evolved BoNT/E protease to cleave phosphatase and tensin homolog (PTEN) but not any natural BoNT protease substrate in neurons. The evolved proteases display large changes in specificity (218- to >11,000,000-fold) and can retain their ability to form holotoxins that self-deliver into primary neurons. These findings establish a versatile platform for reprogramming proteases to selectively cleave new targets of therapeutic interest.

PubMed: 33602850
DOI: 10.1126/science.abf5972

実験手法

X-RAY DIFFRACTION (1.8 Å)