CaltechAUTHORS
  A Caltech Library Service

SCHEMA-Designed Variants of Human Arginase I and II Reveal Sequence Elements Important to Stability and Catalysis

Romero, Philip A. and Stone, Everett and Lamb, Candice and Chantranupong, Lynne and Krause, Andreas and Miklos, Alexandr E. and Hughes, Randall A. and Fechtel, Blake and Ellington, Andrew D. and Arnold, Frances H. and Georgiou, George (2012) SCHEMA-Designed Variants of Human Arginase I and II Reveal Sequence Elements Important to Stability and Catalysis. ACS Synthetic Biology, 1 (6). pp. 221-228. ISSN 2161-5063. PMCID PMC3378063. https://resolver.caltech.edu/CaltechAUTHORS:20120716-102139957

[img] PDF - Accepted Version
See Usage Policy.

815Kb

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20120716-102139957

Abstract

Arginases catalyze the divalent cation-dependent hydrolysis of l-arginine to urea and l-ornithine. There is significant interest in using arginase as a therapeutic anti-neogenic agent against l-arginine auxotrophic tumors and in enzyme replacement therapy for treating hyperargininemia. Both therapeutic applications require enzymes with sufficient stability under physiological conditions. To explore sequence elements that contribute to arginase stability we used SCHEMA-guided recombination to design a library of chimeric enzymes composed of sequence fragments from the two human isozymes Arginase I and II. We then developed a novel active learning algorithm that selects sequences from this library that are both highly informative and functional. Using high-throughput gene synthesis and our two-step active learning algorithm, we were able to rapidly create a small but highly informative set of seven enzymatically active chimeras that had an average variant distance of 40 mutations from the closest parent arginase. Within this set of sequences, linear regression was used to identify the sequence elements that contribute to the long-term stability of human arginase under physiological conditions. This approach revealed a striking correlation between the isoelectric point and the long-term stability of the enzyme to deactivation under physiological conditions.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/sb300014tDOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3378063/PubMed CentralArticle
ORCID:
AuthorORCID
Romero, Philip A.0000-0002-2586-7263
Krause, Andreas0000-0001-7260-9673
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2012 American Chemical Society. Published In Issue June 15, 2012; Article ASAP April 10, 2012; Just Accepted Manuscript March 30, 2012; Received: March 07, 2012. This project was supported by grants (#HF0032 and F-1654) from TI3D/Welch Foundation and National Institutes of Health (CA 139059). In addition, this work was supported by the National Security Science and Engineering Faculty Fellowship (FA9550-10-1-0169), and L.C. was supported by a fellowship from the Arnold & Mabel Beckman Foundation. The authors also acknowledge the National Institutes of Health, ARRA (grant R01-GM068664 to FHA) for funding SCHEMA library design, and the U.S. Army Research Office, Institute for Collaborative Biotechnologies (grant W911NF-09-D-0001 to FHA) for funding the regression analysis work. These contents are solely the responsibility of the authors and do not necessarily represent the official views of the sponsors.
Funders:
Funding AgencyGrant Number
TI3D/Robert A. Welch FoundationHF0032
TI3D/Robert A. Welch FoundationF-1654
NSFCA 139059
National Security Science and Engineering Faculty FellowshipFA9550-10-1-0169
Arnold and Mabel Beckman FoundationUNSPECIFIED
NIHR01-GM068664
Army Research Office (ARO)W911NF-09-D-0001
American Recovery and Reinvestment Act (ARRA)UNSPECIFIED
Subject Keywords:enzyme engineering; arginase; homologous recombination; SCHEMA library design; active learning; protein stability
Issue or Number:6
PubMed Central ID:PMC3378063
Record Number:CaltechAUTHORS:20120716-102139957
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20120716-102139957
Official Citation:SCHEMA-Designed Variants of Human Arginase I and II Reveal Sequence Elements Important to Stability and Catalysis Philip A. Romero, Everett Stone, Candice Lamb, Lynne Chantranupong, Andreas Krause, Aleksandr E. Miklos, Randall A. Hughes, Blake Fechtel, Andrew D. Ellington, Frances H. Arnold, and George Georgiou ACS Synthetic Biology 2012 1 (6), 221-228
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:32465
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:16 Jul 2012 17:56
Last Modified:27 Apr 2020 23:10

Repository Staff Only: item control page