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

Journal Article · Mon Jan 01 04:00:00 EST 2007 · Archives of Biochemistry and Biophysics

DOI:https://doi.org/10.1016/j.abb.2007.04.036· OSTI ID:930015

Di Costanzo, L; Moulin, M; Haertlein, M; Meilleur, F; Christianson, D

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.

• Research Organization: Brookhaven National Laboratory (BNL) National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: AC02-98CH10886
• OSTI ID: 930015
• Report Number(s): BNL--80629-2008-JA
• Journal Information: Archives of Biochemistry and Biophysics, Journal Name: Archives of Biochemistry and Biophysics Journal Issue: 1 Vol. 465; ISSN ABBIA4; ISSN 0003-9861
• Country of Publication: United States
• Language: English

Related Subjects

36 MATERIALS SCIENCE
ARGINASE
COMPLEXES
CRYSTAL STRUCTURE
CRYSTALS
FUNCTIONALS
HUMAN POPULATIONS
HYDROLYSIS
MANGANESE
NEUTRON DIFFRACTION
NEUTRONS
PURIFICATION
UREA
national synchrotron light source