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

doi:10.1016/j.abb.2007.04.036

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

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

Publication Date:: Mon Jan 01 00:00:00 EST 2007

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.

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                    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.

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                    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.

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@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}

}

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DOI: 10.1016/j.abb.2007.04.036

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