Expression, purification, crystallization, and preliminary X-ray analysis of the N-terminal domain of Escherichia coli adenylyl transferase

Yibin Xu; Daying Wen; Paula Clancy; Paul D. Carr; David L. Ollis; Subhash G. Vasudevan

doi:10.1016/j.pep.2003.11.001

Expression, purification, crystallization, and preliminary X-ray analysis of the N-terminal domain of Escherichia coli adenylyl transferase

Yibin Xu^*, Daying Wen, Paula Clancy, Paul D. Carr, David L. Ollis, Subhash G. Vasudevan

^*Corresponding author for this work

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

7 Citations (Scopus)

Abstract

A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domain is truncated at the end of a predicted helix and prior to a Q-linker. The domain was found to be very soluble and stable so that it could be purified to homogeneity and crystallized. This construct has deadenylylation activity that is independent of the low nitrogen status indicator PII-UMP. The crystals belong to space group P3₁21 or its enantiomorph P3₂21 with a = b = 116.6 Å and c = 67.6 Å.

Original language	English
Pages (from-to)	142-146
Number of pages	5
Journal	Protein Expression and Purification
Volume	34
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2003.11.001
Publication status	Published - Mar 2004

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10.1016/j.pep.2003.11.001

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title = "Expression, purification, crystallization, and preliminary X-ray analysis of the N-terminal domain of Escherichia coli adenylyl transferase",

abstract = "A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domain is truncated at the end of a predicted helix and prior to a Q-linker. The domain was found to be very soluble and stable so that it could be purified to homogeneity and crystallized. This construct has deadenylylation activity that is independent of the low nitrogen status indicator PII-UMP. The crystals belong to space group P3121 or its enantiomorph P3221 with a = b = 116.6 {\AA} and c = 67.6 {\AA}.",

keywords = "Bifunctional enzyme, Crystallization, Domain structure, Nitrogen assimilation, Nucleotidyl transferase",

author = "Yibin Xu and Daying Wen and Paula Clancy and Carr, {Paul D.} and Ollis, {David L.} and Vasudevan, {Subhash G.}",

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T1 - Expression, purification, crystallization, and preliminary X-ray analysis of the N-terminal domain of Escherichia coli adenylyl transferase

AU - Xu, Yibin

AU - Wen, Daying

AU - Clancy, Paula

AU - Carr, Paul D.

AU - Ollis, David L.

AU - Vasudevan, Subhash G.

PY - 2004/3

Y1 - 2004/3

N2 - A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domain is truncated at the end of a predicted helix and prior to a Q-linker. The domain was found to be very soluble and stable so that it could be purified to homogeneity and crystallized. This construct has deadenylylation activity that is independent of the low nitrogen status indicator PII-UMP. The crystals belong to space group P3121 or its enantiomorph P3221 with a = b = 116.6 Å and c = 67.6 Å.

AB - A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domain is truncated at the end of a predicted helix and prior to a Q-linker. The domain was found to be very soluble and stable so that it could be purified to homogeneity and crystallized. This construct has deadenylylation activity that is independent of the low nitrogen status indicator PII-UMP. The crystals belong to space group P3121 or its enantiomorph P3221 with a = b = 116.6 Å and c = 67.6 Å.

KW - Bifunctional enzyme

KW - Crystallization

KW - Domain structure

KW - Nitrogen assimilation

KW - Nucleotidyl transferase

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U2 - 10.1016/j.pep.2003.11.001

DO - 10.1016/j.pep.2003.11.001

M3 - Article

SN - 1046-5928

VL - 34

SP - 142

EP - 146

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 1

ER -

Expression, purification, crystallization, and preliminary X-ray analysis of the N-terminal domain of Escherichia coli adenylyl transferase

Abstract

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