Enzyme Cascade with Horseradish Peroxidase Readout for High-Throughput Screening and Engineering of Human Arginase-1

被引:6
作者
De Santaella, Jaime Fernandez [1 ,2 ,3 ]
Ren, Jin [1 ,2 ]
Vanella, Rosario [1 ,2 ]
Nash, Michael A. [1 ,2 ,3 ,4 ]
机构
[1] Univ Basel, Inst Phys Chem, Dept Chem, CH-4058 Basel, Switzerland
[2] Dept Biosyst Sci & Engn, ETH Zurich, CH-4058 Basel, Switzerland
[3] Natl Ctr Competence Res NCCR, Mol Syst Engn, CH-4058 Basel, Switzerland
[4] Swiss Nanosci Inst, CH-4056 Basel, Switzerland
基金
瑞士国家科学基金会;
关键词
CRYSTAL-STRUCTURE; RATIONAL DESIGN; LIVER ARGINASE; INHIBITION; EVOLUTION; ASSAY;
D O I
10.1021/acs.analchem.2c05429
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We report an enzyme cascade with horseradish peroxidase-based readout for screening human arginase-1 (hArg1) activity. We combined the four enzymes hArg1, ornithine decarboxylase, putrescine oxidase, and horseradish peroxidase in a reaction cascade that generated colorimetric or fluorescent signals in response to hArg1 activity and used this cascade to assay wild-type and variant hArg1 sequences as soluble enzymes and displayed on the surface of Escherichia coli. We screened a curated 13-member hArg1 library covering mutations that modified the electrostatic environment surrounding catalytic residues D128 and H141, and identified the R21E variant with a 13% enhanced catalytic turnover rate compared to wild type. Our scalable one-pot single-step arginase assay with continuous kinetic readout is amenable to high-throughput screening and directed evolution of arginase libraries and testing drug candidates for arginase inhibition.
引用
收藏
页码:7150 / 7157
页数:8
相关论文
共 46 条
[1]   Mutational analysis of substrate recognition by human arginase type I -: agmatinase activity of the N130D variant [J].
Alarcon, Ricardo ;
Orellana, Maria S. ;
Neira, Benita ;
Uribe, Elena ;
Garcia, Jose R. ;
Carvajal, Nelson .
FEBS JOURNAL, 2006, 273 (24) :5625-5631
[2]   Identification of an Arginase II Inhibitor via RapidFire Mass Spectrometry Combined with Hydrophilic Interaction Chromatography [J].
Asano, Wataru ;
Takahashi, Yu ;
Kawano, Motoaki ;
Hantani, Yoshiji .
SLAS DISCOVERY, 2019, 24 (04) :457-465
[3]   A SENSITIVE METHOD FOR COLORIMETRIC DETERMINATION OF UREA [J].
BEALE, RN ;
CROFT, D .
JOURNAL OF CLINICAL PATHOLOGY, 1961, 14 (04) :418-&
[4]   Multistep enzyme cascades as a route towards green and sustainable pharmaceutical syntheses [J].
Benitez-Mateos, Ana, I ;
Padrosa, David Roura ;
Paradisi, Francesca .
NATURE CHEMISTRY, 2022, 14 (05) :489-499
[5]  
Biotherapeutics, 2019, PHASE 12 STUDY AEB11
[6]   IMMOBILIZED ENZYME ELECTRODE FOR DETERMINATION OF ARGINASE [J].
BOOKER, HE ;
HASLAM, JL .
ANALYTICAL CHEMISTRY, 1974, 46 (08) :1054-1060
[7]   Chemical modification and site-directed mutagenesis of human liver arginase:: Evidence that the imidazole group of histidine-141 is not involved in substrate binding [J].
Carvajal, N ;
Olate, J ;
Salas, M ;
Uribe, E ;
López, V ;
Herrera, P ;
Cerpa, J .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1999, 371 (02) :202-206
[8]   A phase II clinical study on the efficacy and predictive biomarker of pegylated recombinant arginase on hepatocellular carcinoma [J].
Chan, Stephen L. ;
Cheng, Paul N. M. ;
Liu, Angela M. ;
Chan, Landon L. ;
Li, Leung ;
Chu, Cheuk M. ;
Chong, Charing C. N. ;
Lau, Yat M. ;
Yeo, Winnie ;
Ng, Kelvin K. C. ;
Yu, Simon C. H. ;
Mok, Tony S. K. ;
Chan, Anthony W. H. .
INVESTIGATIONAL NEW DRUGS, 2021, 39 (05) :1375-1382
[9]   Rational design of ornithine decarboxylase with high catalytic activity for the production of putrescine [J].
Choi, Hyang ;
Kyeong, Hyun-Ho ;
Choi, Jung Min ;
Kim, Hak-Sung .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2014, 98 (17) :7483-7490
[10]   Arginase as a Potential Biomarker of Disease Progression: A Molecular Imaging Perspective [J].
Clemente, Goncalo S. ;
van Waarde, Aren ;
Antunes, Ines F. ;
Domling, Alexander ;
Elsinga, Philip H. .
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2020, 21 (15) :1-36