7REP

Crystal structure of an engineered variant of single-chain Penicillin G Acylase from Kluyvera cryocrescens (A1-Ac Rd3CHis)

7REP の概要

• エントリーDOI: 10.2210/pdb7rep/pdb
• 関連するPDBエントリー: 7REO
• 分子名称: Penicillin G acylase, CALCIUM ION, phenylmethanesulfonic acid, ... (4 entities in total)
• 機能のキーワード: penicillin, acylase, hydrolase
• 由来する生物種: Kluyvera cryocrescens
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 86471.39

構造登録者

Orth, P. (登録日: 2021-07-13, 公開日: 2021-11-17, 最終更新日: 2024-10-30)

主引用文献

Fryszkowska, A.,An, C.,Alvizo, O.,Banerjee, G.,Canada, K.A.,Cao, Y.,DeMong, D.,Devine, P.N.,Duan, D.,Elgart, D.M.,Farasat, I.,Gauthier, D.R.,Guidry, E.N.,Jia, X.,Kong, J.,Kruse, N.,Lexa, K.W.,Makarov, A.A.,Mann, B.F.,Milczek, E.M.,Mitchell, V.,Nazor, J.,Neri, C.,Orr, R.K.,Orth, P.,Phillips, E.M.,Riggins, J.N.,Schafer, W.A.,Silverman, S.M.,Strulson, C.A.,Subramanian, N.,Voladri, R.,Yang, H.,Yang, J.,Yi, X.,Zhang, X.,Zhong, W.
A chemoenzymatic strategy for site-selective functionalization of native peptides and proteins.
Science, 376:1321-1327, 2022

PubMed Abstract: The emergence of new therapeutic modalities requires complementary tools for their efficient syntheses. Availability of methodologies for site-selective modification of biomolecules remains a long-standing challenge, given the inherent complexity and the presence of repeating residues that bear functional groups with similar reactivity profiles. We describe a bioconjugation strategy for modification of native peptides relying on high site selectivity conveyed by enzymes. We engineered penicillin G acylases to distinguish among free amino moieties of insulin (two at amino termini and an internal lysine) and manipulate cleavable phenylacetamide groups in a programmable manner to form protected insulin derivatives. This enables selective and specific chemical ligation to synthesize homogeneous bioconjugates, improving yield and purity compared to the existing methods, and generally opens avenues in the functionalization of native proteins to access biological probes or drugs.

PubMed: 35709255
DOI: 10.1126/science.abn2009

実験手法

X-RAY DIFFRACTION (2.192 Å)