Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1

Li Ying Chen; Pei Hsun Wu; Hong Hsiang Guan; Hoong Kun Fun; Hwan You Chang; Chun Jung Chen

doi:10.1107/S2053230X15004197

Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1

Li Ying Chen
, Pei Hsun Wu
, Hong Hsiang Guan
, Hoong Kun Fun
, Hwan You Chang
, Chun Jung Chen^*

^*Corresponding author for this work

National Cheng Kung University
National Synchrotron Radiation Research Center Taiwan
National Tsing Hua University
King Saud University
Universiti Sains Malaysia

Research output: Contribution to journal › Journal Article › peer-review

Abstract

The phosphatase domain (PA3346PD) of the response regulator PA3346 modulates the downstream anti-anti-σ factor PA3347 to regulate swarming motility in Pseudomonas aeruginosa PAO1. PA3346PD, which comprises the protein phosphatase 2C domain (PP2C), is classified as a Ser/Thr phosphatase of the Mg²⁺-or Mn²⁺-dependent protein phosphatase (PPM) family. The recombinant PA3346PD, with molecular mass 26 kDa, was overexpressed in Escherichia coli, purified on an Ni²⁺-NTA agarose column and crystallized by the sitting-drop vapour-diffusion method. X-ray diffraction data were collected from PA3346PD crystals to a resolution of 2.58 Å and the crystals belonged to space group I4132 or I4332, with unit-cell parameter a = 157.61 Å. Preliminary analysis indicates the presence of a monomer of PA3346PD in the asymmetric unit with a solvent content of 58.4%.

Original language	English
Pages (from-to)	434-437
Number of pages	4
Journal	Acta Crystallographica Section F: Structural Biology Communications
Volume	71
DOIs	https://doi.org/10.1107/S2053230X15004197
State	Published - 01 04 2015
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2015 International Union of Crystallography.

Keywords

Pseudomonas aeruginosa
phosphatase domain
response regulator

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10.1107/S2053230X15004197

Cite this

Chen, L. Y., Wu, P. H., Guan, H. H., Fun, H. K., Chang, H. Y., & Chen, C. J. (2015). Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1. Acta Crystallographica Section F: Structural Biology Communications, 71, 434-437. https://doi.org/10.1107/S2053230X15004197

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title = "Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1",

abstract = "The phosphatase domain (PA3346PD) of the response regulator PA3346 modulates the downstream anti-anti-σ factor PA3347 to regulate swarming motility in Pseudomonas aeruginosa PAO1. PA3346PD, which comprises the protein phosphatase 2C domain (PP2C), is classified as a Ser/Thr phosphatase of the Mg2+-or Mn2+-dependent protein phosphatase (PPM) family. The recombinant PA3346PD, with molecular mass 26 kDa, was overexpressed in Escherichia coli, purified on an Ni2+-NTA agarose column and crystallized by the sitting-drop vapour-diffusion method. X-ray diffraction data were collected from PA3346PD crystals to a resolution of 2.58 {\AA} and the crystals belonged to space group I4132 or I4332, with unit-cell parameter a = 157.61 {\AA}. Preliminary analysis indicates the presence of a monomer of PA3346PD in the asymmetric unit with a solvent content of 58.4\%.",

keywords = "Pseudomonas aeruginosa, phosphatase domain, response regulator",

author = "Chen, \{Li Ying\} and Wu, \{Pei Hsun\} and Guan, \{Hong Hsiang\} and Fun, \{Hoong Kun\} and Chang, \{Hwan You\} and Chen, \{Chun Jung\}",

note = "Publisher Copyright: {\textcopyright} 2015 International Union of Crystallography.",

year = "2015",

month = apr,

day = "1",

doi = "10.1107/S2053230X15004197",

language = "英语",

volume = "71",

pages = "434--437",

journal = "Acta Crystallographica Section F: Structural Biology Communications",

issn = "2053-230X",

publisher = "John Wiley and Sons Ltd",

}

Chen, LY, Wu, PH, Guan, HH, Fun, HK, Chang, HY & Chen, CJ 2015, 'Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1', Acta Crystallographica Section F: Structural Biology Communications, vol. 71, pp. 434-437. https://doi.org/10.1107/S2053230X15004197

Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1. / Chen, Li Ying; Wu, Pei Hsun; Guan, Hong Hsiang et al.
In: Acta Crystallographica Section F: Structural Biology Communications, Vol. 71, 01.04.2015, p. 434-437.

Research output: Contribution to journal › Journal Article › peer-review

TY - JOUR

T1 - Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1

AU - Chen, Li Ying

AU - Wu, Pei Hsun

AU - Guan, Hong Hsiang

AU - Fun, Hoong Kun

AU - Chang, Hwan You

AU - Chen, Chun Jung

PY - 2015/4/1

Y1 - 2015/4/1

N2 - The phosphatase domain (PA3346PD) of the response regulator PA3346 modulates the downstream anti-anti-σ factor PA3347 to regulate swarming motility in Pseudomonas aeruginosa PAO1. PA3346PD, which comprises the protein phosphatase 2C domain (PP2C), is classified as a Ser/Thr phosphatase of the Mg2+-or Mn2+-dependent protein phosphatase (PPM) family. The recombinant PA3346PD, with molecular mass 26 kDa, was overexpressed in Escherichia coli, purified on an Ni2+-NTA agarose column and crystallized by the sitting-drop vapour-diffusion method. X-ray diffraction data were collected from PA3346PD crystals to a resolution of 2.58 Å and the crystals belonged to space group I4132 or I4332, with unit-cell parameter a = 157.61 Å. Preliminary analysis indicates the presence of a monomer of PA3346PD in the asymmetric unit with a solvent content of 58.4%.

AB - The phosphatase domain (PA3346PD) of the response regulator PA3346 modulates the downstream anti-anti-σ factor PA3347 to regulate swarming motility in Pseudomonas aeruginosa PAO1. PA3346PD, which comprises the protein phosphatase 2C domain (PP2C), is classified as a Ser/Thr phosphatase of the Mg2+-or Mn2+-dependent protein phosphatase (PPM) family. The recombinant PA3346PD, with molecular mass 26 kDa, was overexpressed in Escherichia coli, purified on an Ni2+-NTA agarose column and crystallized by the sitting-drop vapour-diffusion method. X-ray diffraction data were collected from PA3346PD crystals to a resolution of 2.58 Å and the crystals belonged to space group I4132 or I4332, with unit-cell parameter a = 157.61 Å. Preliminary analysis indicates the presence of a monomer of PA3346PD in the asymmetric unit with a solvent content of 58.4%.

KW - Pseudomonas aeruginosa

KW - phosphatase domain

KW - response regulator

UR - https://www.scopus.com/pages/publications/84927622137

U2 - 10.1107/S2053230X15004197

DO - 10.1107/S2053230X15004197

M3 - 文章

C2 - 25849505

AN - SCOPUS:84927622137

SN - 2053-230X

VL - 71

SP - 434

EP - 437

JO - Acta Crystallographica Section F: Structural Biology Communications

JF - Acta Crystallographica Section F: Structural Biology Communications

ER -

Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1

Abstract

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Keywords

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