Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase

Jaldappagari Seetharamappa; Muse Oke; Huanting Liu; Stephen A McMahon; Kenneth A Johnson; Lester Carter; Mark Dorward; Michal Zawadzki; Ian M Overton; C A Johannes van Niekirk; Shirley Graham; Catherine H Botting; Garry L Taylor; Malcolm F White; Geoffrey J Barton; Peter J Coote; James H Naismith

doi:10.1107/S1744309107019562

Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase

Jaldappagari Seetharamappa
, Muse Oke
, Huanting Liu
, Stephen A McMahon
, Kenneth A Johnson
, Lester Carter
, Mark Dorward
, Michal Zawadzki
, Ian M Overton
, C A Johannes van Niekirk
, Shirley Graham
, Catherine H Botting
, Garry L Taylor
, Malcolm F White
, Geoffrey J Barton
, Peter J Coote
, James H Naismith

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

4 Citations (Scopus)

Abstract

Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared with a methicillin-sensitive strain. The protein was expressed in Escherichia coli, purified and crystallized. The protein crystallized in a primitive orthorhombic Laue group with unit-cell parameters a = 83.6, b = 91.3, c = 106.0 A, alpha = beta = gamma = 90 degrees. Analysis of the systematic absences along the three principal axes indicated the space group to be P2(1)2(1)2(1). A complete data set was collected to 2.5 A resolution.

Original language	English
Pages (from-to)	452-6
Number of pages	5
Journal	Acta Crystallographica F-Structural Biology and Crystallization Communications
Volume	63
Issue number	Pt 5
DOIs	https://doi.org/10.1107/S1744309107019562
Publication status	Published - 1 May 2007

Keywords

Bacterial Proteins
Chromatography, Affinity
Cloning, Molecular
Crystallization
Methicillin Resistance
Protein Conformation
Staphylococcus aureus
Transaminases

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10.1107/S1744309107019562

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Seetharamappa, J., Oke, M., Liu, H., McMahon, S. A., Johnson, K. A., Carter, L., Dorward, M., Zawadzki, M., Overton, I. M., van Niekirk, C. A. J., Graham, S., Botting, C. H., Taylor, G. L., White, M. F., Barton, G. J., Coote, P. J., & Naismith, J. H. (2007). Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase. Acta Crystallographica F-Structural Biology and Crystallization Communications, 63(Pt 5), 452-6. https://doi.org/10.1107/S1744309107019562

Seetharamappa, Jaldappagari ; Oke, Muse ; Liu, Huanting et al. / Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase. In: Acta Crystallographica F-Structural Biology and Crystallization Communications. 2007 ; Vol. 63, No. Pt 5. pp. 452-6.

@article{f30963acb83a4ee09729e8c91f51c378,

title = "Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase",

abstract = "Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared with a methicillin-sensitive strain. The protein was expressed in Escherichia coli, purified and crystallized. The protein crystallized in a primitive orthorhombic Laue group with unit-cell parameters a = 83.6, b = 91.3, c = 106.0 A, alpha = beta = gamma = 90 degrees. Analysis of the systematic absences along the three principal axes indicated the space group to be P2(1)2(1)2(1). A complete data set was collected to 2.5 A resolution.",

keywords = "Bacterial Proteins, Chromatography, Affinity, Cloning, Molecular, Crystallization, Methicillin Resistance, Protein Conformation, Staphylococcus aureus, Transaminases",

author = "Jaldappagari Seetharamappa and Muse Oke and Huanting Liu and McMahon, \{Stephen A\} and Johnson, \{Kenneth A\} and Lester Carter and Mark Dorward and Michal Zawadzki and Overton, \{Ian M\} and \{van Niekirk\}, \{C A Johannes\} and Shirley Graham and Botting, \{Catherine H\} and Taylor, \{Garry L\} and White, \{Malcolm F\} and Barton, \{Geoffrey J\} and Coote, \{Peter J\} and Naismith, \{James H\}",

year = "2007",

month = may,

day = "1",

doi = "10.1107/S1744309107019562",

language = "English",

volume = "63",

pages = "452--6",

journal = "Acta Crystallographica F-Structural Biology and Crystallization Communications",

issn = "1744-3091",

publisher = "International Union of Crystallography",

number = "Pt 5",

}

Seetharamappa, J, Oke, M, Liu, H, McMahon, SA, Johnson, KA, Carter, L, Dorward, M, Zawadzki, M, Overton, IM, van Niekirk, CAJ, Graham, S, Botting, CH, Taylor, GL, White, MF, Barton, GJ, Coote, PJ & Naismith, JH 2007, 'Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase', Acta Crystallographica F-Structural Biology and Crystallization Communications, vol. 63, no. Pt 5, pp. 452-6. https://doi.org/10.1107/S1744309107019562

Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase. / Seetharamappa, Jaldappagari; Oke, Muse; Liu, Huanting et al.
In: Acta Crystallographica F-Structural Biology and Crystallization Communications, Vol. 63, No. Pt 5, 01.05.2007, p. 452-6.

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

TY - JOUR

T1 - Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase

AU - Seetharamappa, Jaldappagari

AU - Oke, Muse

AU - Liu, Huanting

AU - McMahon, Stephen A

AU - Johnson, Kenneth A

AU - Carter, Lester

AU - Dorward, Mark

AU - Zawadzki, Michal

AU - Overton, Ian M

AU - van Niekirk, C A Johannes

AU - Graham, Shirley

AU - Botting, Catherine H

AU - Taylor, Garry L

AU - White, Malcolm F

AU - Barton, Geoffrey J

AU - Coote, Peter J

AU - Naismith, James H

PY - 2007/5/1

Y1 - 2007/5/1

N2 - Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared with a methicillin-sensitive strain. The protein was expressed in Escherichia coli, purified and crystallized. The protein crystallized in a primitive orthorhombic Laue group with unit-cell parameters a = 83.6, b = 91.3, c = 106.0 A, alpha = beta = gamma = 90 degrees. Analysis of the systematic absences along the three principal axes indicated the space group to be P2(1)2(1)2(1). A complete data set was collected to 2.5 A resolution.

AB - Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase with a molecular weight of 48,168 Da, was overexpressed in methicillin-resistant Staphylococcus aureus compared with a methicillin-sensitive strain. The protein was expressed in Escherichia coli, purified and crystallized. The protein crystallized in a primitive orthorhombic Laue group with unit-cell parameters a = 83.6, b = 91.3, c = 106.0 A, alpha = beta = gamma = 90 degrees. Analysis of the systematic absences along the three principal axes indicated the space group to be P2(1)2(1)2(1). A complete data set was collected to 2.5 A resolution.

KW - Bacterial Proteins

KW - Chromatography, Affinity

KW - Cloning, Molecular

KW - Crystallization

KW - Methicillin Resistance

KW - Protein Conformation

KW - Staphylococcus aureus

KW - Transaminases

U2 - 10.1107/S1744309107019562

DO - 10.1107/S1744309107019562

M3 - Article

C2 - 17565195

SN - 1744-3091

VL - 63

SP - 452

EP - 456

JO - Acta Crystallographica F-Structural Biology and Crystallization Communications

JF - Acta Crystallographica F-Structural Biology and Crystallization Communications

IS - Pt 5

ER -

Seetharamappa J, Oke M, Liu H, McMahon SA, Johnson KA, Carter L et al. Expression, purification, crystallization, data collection and preliminary biochemical characterization of methicillin-resistant Staphylococcus aureus Sar2028, an aspartate/tyrosine/phenylalanine pyridoxal-5'-phosphate-dependent aminotransferase. Acta Crystallographica F-Structural Biology and Crystallization Communications. 2007 May 1;63(Pt 5):452-6. doi: 10.1107/S1744309107019562