Acrylate-derived RAFT Polymers for Enzyme Hyperactivation - Boosting the α-Chymotrypsin Enzyme Activity Using Tailor-Made Poly(2-Carboxyethyl)acrylate (PCEA)

被引：0

作者：

Muellers, Yannic ^{[1
,2
]}

Sadr, Ahmad Shahir ^{[3
]}

Schenderlein, Matthias ^{[2
]}

Pallab, Nazim ^{[1
,2
]}

Davari, Mehdi D. ^{[3
]}

Glebe, Ulrich ^{[1
,2
]}

Reifarth, Martin ^{[1
,2
]}

机构：

[1] Univ Potsdam, Inst Chem, Karl Liebknecht Str 24-25, D-14476 Potsdam, Germany

[2] Fraunhofer Inst Appl Polymer Res IAP, Geiselbergstr 69, D-14476 Potsdam, Germany

[3] Leibniz Inst Plant Biochem, Dept Bioorgan Chem, Weinberg 3, D-06120 Halle, Germany

来源：

CHEMCATCHEM | 2024年 / 16卷 / 11期

关键词：

Enzyme hyperactivation; Polyacrylate; RAFT polymerization; Polymer/enzyme complexes; Molecular dynamics simulation; PARAMETERIZATION; STABILITY; STRATEGY; PROTEIN; PREDICT; VIEW;

D O I：

10.1002/cctc.202301685

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

We study the hyperactivation of alpha-chymotrypsin (alpha-ChT) using the acrylate polymer poly(2-carboxyethyl) acrylate (PCEA) in comparison to the commonly used poly(acrylic acid) (PAA). The polymers are added during the enzymatic cleavage reaction of the substrate N-glycyl-L-phenylalanine p-nitroanilide (GPNA). Enzyme activity assays reveal a pronounced enzyme hyperactivation capacity of PCEA, which reaches up to 950 % activity enhancement, and is significantly enhanced to PAA (revealing an activity enhancement of approx. 450 %). In a combined experimental and computational study, we investigate alpha-ChT/polymer interactions to elucidate the hyperactivation mechanism of the enzyme. Isothermal titration calorimetry reveals a pronounced complexation between the polymer and the enzyme. Docking simulations reveal that binding of polymers significantly improves the binding affinity of GPNA to alpha-ChT. Notably, a higher binding affinity is found for the alpha-ChT/PCEA compared to the alpha-ChT/PAA complex. Further molecular dynamics (MD) simulations reveal changes in the size of the active site in the enzyme/polymer complexes, with PCEA inducing a more pronounced alteration compared to PAA, facilitating an easier access for the substrate to the active site of alpha-ChT. We study the hyperactivation of alpha-chymotrypsin (alpha-ChT) here, using acrylate-derived PCEA polymer. Enzyme activity assays revealed a pronounced enzyme hyperactivation capacity being superior to widespread PAA polymers. In a combined experimental and computational study, we investigate alpha-ChT/polymer interactions to elucidate the hyperactivation mechanism.+ image

引用

页数：7