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Title: Expression, Purification, Assay, and Crystal Structure of Perdeuterated Human Arginase I

Abstract

Arginase is a manganese metalloenzyme that catalyzes the hydrolysis of L-arginine to yield L-ornithine and urea. In order to establish a foundation for future neutron diffraction studies that will provide conclusive structural information regarding proton/deuteron positions in enzyme-inhibitor complexes, we have expressed, purified, assayed, and determined the X-ray crystal structure of perdeuterated (i.e., fully deuterated) human arginase I complexed with 2(S)-amino-6-boronohexanoic acid (ABH) at 1.90 {angstrom} resolution. Prior to the neutron diffraction experiment, it is important to establish that perdeuteration does not cause any unanticipated structural or functional changes. Accordingly, we find that perdeuterated human arginase I exhibits catalytic activity essentially identical to that of the unlabeled enzyme. Additionally, the structure of the perdeuterated human arginase I-ABH complex is identical to that of the corresponding complex with the unlabeled enzyme. Therefore, we conclude that crystals of the perdeuterated human arginase I-ABH complex are suitable for neutron crystallographic study.

Authors:
; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
930015
Report Number(s):
BNL-80629-2008-JA
Journal ID: ISSN 0003-9861; ABBIA4; TRN: US200822%%1166
DOE Contract Number:
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Archives of Biochemistry and Biophysics; Journal Volume: 465; Journal Issue: 1
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; ARGINASE; COMPLEXES; CRYSTAL STRUCTURE; CRYSTALS; FUNCTIONALS; HUMAN POPULATIONS; HYDROLYSIS; MANGANESE; NEUTRON DIFFRACTION; NEUTRONS; PURIFICATION; UREA; national synchrotron light source

Citation Formats

Di Costanzo,L., Moulin, M., Haertlein, M., Meilleur, F., and Christianson, D. Expression, Purification, Assay, and Crystal Structure of Perdeuterated Human Arginase I. United States: N. p., 2007. Web. doi:10.1016/j.abb.2007.04.036.
Di Costanzo,L., Moulin, M., Haertlein, M., Meilleur, F., & Christianson, D. Expression, Purification, Assay, and Crystal Structure of Perdeuterated Human Arginase I. United States. doi:10.1016/j.abb.2007.04.036.
Di Costanzo,L., Moulin, M., Haertlein, M., Meilleur, F., and Christianson, D. Mon . "Expression, Purification, Assay, and Crystal Structure of Perdeuterated Human Arginase I". United States. doi:10.1016/j.abb.2007.04.036.
@article{osti_930015,
title = {Expression, Purification, Assay, and Crystal Structure of Perdeuterated Human Arginase I},
author = {Di Costanzo,L. and Moulin, M. and Haertlein, M. and Meilleur, F. and Christianson, D.},
abstractNote = {Arginase is a manganese metalloenzyme that catalyzes the hydrolysis of L-arginine to yield L-ornithine and urea. In order to establish a foundation for future neutron diffraction studies that will provide conclusive structural information regarding proton/deuteron positions in enzyme-inhibitor complexes, we have expressed, purified, assayed, and determined the X-ray crystal structure of perdeuterated (i.e., fully deuterated) human arginase I complexed with 2(S)-amino-6-boronohexanoic acid (ABH) at 1.90 {angstrom} resolution. Prior to the neutron diffraction experiment, it is important to establish that perdeuteration does not cause any unanticipated structural or functional changes. Accordingly, we find that perdeuterated human arginase I exhibits catalytic activity essentially identical to that of the unlabeled enzyme. Additionally, the structure of the perdeuterated human arginase I-ABH complex is identical to that of the corresponding complex with the unlabeled enzyme. Therefore, we conclude that crystals of the perdeuterated human arginase I-ABH complex are suitable for neutron crystallographic study.},
doi = {10.1016/j.abb.2007.04.036},
journal = {Archives of Biochemistry and Biophysics},
number = 1,
volume = 465,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}