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Title: Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution

Abstract

The enzyme dihydrodipicolinate synthase catalyzes the committed step in the synthesis of diaminopimelate and lysine to facilitate peptidoglycan and protein synthesis. Dihydrodipicolinate synthase catalyzes the condensation of L-aspartate 4-semialdehyde and pyruvate to synthesize L-2,3-dihydrodipicolinate. Here, the cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the pathogenic bacteriumBartonella henselae, the causative bacterium of cat-scratch disease, are presented. Protein crystals were grown in conditions consisting of 20%(w/v) PEG 4000, 100 mMsodium citrate tribasic pH 5.5 and were shown to diffract to ~2.10 Å resolution. They belonged to space groupP2 12 12 1, with unit-cell parametersa= 79.96,b= 106.33,c= 136.25 Å. The finalRvalues wereR r.i.m.= 0.098,R work= 0.183,R free= 0.233.

Authors:
 [1];  [2];  [3];  [2];  [4];  [5];  [1]
  1. Rochester Inst. of Technology, Rochester, NY (United States). Thomas H. Gosnell School of Life Sciences
  2. Seattle Structural Genomics Center for Infectious Disease (United States); Center for Infectious Disease Research, Seattle, WA (United States)
  3. Univ. of Canterbury, Christchurch (New Zealand). Biomolecular Interaction Centre, School of Biological Sciences
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Berkeley Center for Structural Biology
  5. Seattle Structural Genomics Center for Infectious Disease (United States); Center for Infectious Disease Research, Seattle, WA (United States); Univ. of Washington, Seattle, WA (United States). Dept. of Global Health; Univ. of Washington, Seattle, WA (United States). Dept. of Biomedical Informatics and Health Education
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Institutes of Health (NIH)
OSTI Identifier:
1378764
DOE Contract Number:  
AC02-05CH11231; HHSN272200700057C; HHSN272201200025C
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F, Structural Biology Communications
Additional Journal Information:
Journal Volume: 72; Journal Issue: 1; Journal ID: ISSN 2053-230X
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Bartonella henselae; dihydrodipicolinate synthase; diaminopimelate; lysine biosynthesis; cat-scratch disease

Citation Formats

Naqvi, Kubra F., Staker, Bart L., Dobson, Renwick C. J., Serbzhinskiy, Dmitry, Sankaran, Banumathi, Myler, Peter J., and Hudson, André O. Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution. United States: N. p., 2016. Web. doi:10.1107/S2053230X15023213.
Naqvi, Kubra F., Staker, Bart L., Dobson, Renwick C. J., Serbzhinskiy, Dmitry, Sankaran, Banumathi, Myler, Peter J., & Hudson, André O. Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution. United States. doi:10.1107/S2053230X15023213.
Naqvi, Kubra F., Staker, Bart L., Dobson, Renwick C. J., Serbzhinskiy, Dmitry, Sankaran, Banumathi, Myler, Peter J., and Hudson, André O. Fri . "Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution". United States. doi:10.1107/S2053230X15023213. https://www.osti.gov/servlets/purl/1378764.
@article{osti_1378764,
title = {Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution},
author = {Naqvi, Kubra F. and Staker, Bart L. and Dobson, Renwick C. J. and Serbzhinskiy, Dmitry and Sankaran, Banumathi and Myler, Peter J. and Hudson, André O.},
abstractNote = {The enzyme dihydrodipicolinate synthase catalyzes the committed step in the synthesis of diaminopimelate and lysine to facilitate peptidoglycan and protein synthesis. Dihydrodipicolinate synthase catalyzes the condensation of L-aspartate 4-semialdehyde and pyruvate to synthesize L-2,3-dihydrodipicolinate. Here, the cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the pathogenic bacteriumBartonella henselae, the causative bacterium of cat-scratch disease, are presented. Protein crystals were grown in conditions consisting of 20%(w/v) PEG 4000, 100 mMsodium citrate tribasic pH 5.5 and were shown to diffract to ~2.10 Å resolution. They belonged to space groupP212121, with unit-cell parametersa= 79.96,b= 106.33,c= 136.25 Å. The finalRvalues wereRr.i.m.= 0.098,Rwork= 0.183,Rfree= 0.233.},
doi = {10.1107/S2053230X15023213},
journal = {Acta Crystallographica. Section F, Structural Biology Communications},
issn = {2053-230X},
number = 1,
volume = 72,
place = {United States},
year = {2016},
month = {1}
}

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