Structural characterization of Escherichia coli sialic acid synthase

Tzann Shun Hwang; Chih Hung Hung; Chin Fen Teo; Guan Ting Chen; Lee Shang Chang; Sung Fang Chen; Yu Ju Chen; Chun Hung Lin

doi:10.1016/S0006-291X(02)00620-4

Structural characterization of Escherichia coli sialic acid synthase

Tzann Shun Hwang, Chih Hung Hung, Chin Fen Teo, Guan Ting Chen, Lee Shang Chang, Sung Fang Chen, Yu Ju Chen, Chun Hung Lin

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

15 Citations (Scopus)

Abstract

Sialic acid synthase encoded by the neuB gene of Escherichia coli catalyzes the condensation of N-acetylmannosamine and phosphoenolpyruvate to form N-acetylneuraminic acid. This report demonstrates the first structural information on sialic acid synthase by CD MALDI-TOF and chemical cross-linking studies. Also a specific cleavage by endogenous protease(s) has been identified at Lys²⁸⁰ of the enzyme (40 kDa) by LC-MS and N-terminal sequencing analyses. The cleavage results in the formation of two inactive fragments of 33 and 7 kDa. The structural analysis indicates that the fragmentation is associated with a significant change of the enzyme from a tetrameric to trimeric form and alterations in both secondary and native quaternary structures.

Original language	English
Pages (from-to)	167-173
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	295
Issue number	1
DOIs	https://doi.org/10.1016/S0006-291X(02)00620-4
Publication status	Published - 2002
Externally published	Yes

Keywords

Circular dichroism (CD)
Matrix-assisted laser desorption (MALDI)/ionization-time-of-flight (TOF)
N-Acetylneuraminic acid (NeuAc)
Sialic acid synthase
Subunit cross-linking

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/S0006-291X(02)00620-4

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title = "Structural characterization of Escherichia coli sialic acid synthase",

abstract = "Sialic acid synthase encoded by the neuB gene of Escherichia coli catalyzes the condensation of N-acetylmannosamine and phosphoenolpyruvate to form N-acetylneuraminic acid. This report demonstrates the first structural information on sialic acid synthase by CD MALDI-TOF and chemical cross-linking studies. Also a specific cleavage by endogenous protease(s) has been identified at Lys280 of the enzyme (40 kDa) by LC-MS and N-terminal sequencing analyses. The cleavage results in the formation of two inactive fragments of 33 and 7 kDa. The structural analysis indicates that the fragmentation is associated with a significant change of the enzyme from a tetrameric to trimeric form and alterations in both secondary and native quaternary structures.",

keywords = "Circular dichroism (CD), Matrix-assisted laser desorption (MALDI)/ionization-time-of-flight (TOF), N-Acetylneuraminic acid (NeuAc), Sialic acid synthase, Subunit cross-linking",

author = "Hwang, {Tzann Shun} and Hung, {Chih Hung} and Teo, {Chin Fen} and Chen, {Guan Ting} and Chang, {Lee Shang} and Chen, {Sung Fang} and Chen, {Yu Ju} and Lin, {Chun Hung}",

note = "Funding Information: The work was supported by National Science Council (NSC-90-2113-M-001-028), National Health Research Institute (NSC-90-2323-B-001-004), and Academia Sinica (Taipei, Taiwan). We are indebted to Prof. Shyh-Horng Chiou at the same institute for his critical reading of the manuscript and suggestions of CD and cross-linking experiments. We also thank Mr. Chuan-Fa Chang for graphic assistance.",

year = "2002",

doi = "10.1016/S0006-291X(02)00620-4",

language = "English",

volume = "295",

pages = "167--173",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Academic Press Inc.",

number = "1",

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TY - JOUR

T1 - Structural characterization of Escherichia coli sialic acid synthase

AU - Hwang, Tzann Shun

AU - Hung, Chih Hung

AU - Teo, Chin Fen

AU - Chen, Guan Ting

AU - Chang, Lee Shang

AU - Chen, Sung Fang

AU - Chen, Yu Ju

AU - Lin, Chun Hung

N1 - Funding Information: The work was supported by National Science Council (NSC-90-2113-M-001-028), National Health Research Institute (NSC-90-2323-B-001-004), and Academia Sinica (Taipei, Taiwan). We are indebted to Prof. Shyh-Horng Chiou at the same institute for his critical reading of the manuscript and suggestions of CD and cross-linking experiments. We also thank Mr. Chuan-Fa Chang for graphic assistance.

PY - 2002

Y1 - 2002

N2 - Sialic acid synthase encoded by the neuB gene of Escherichia coli catalyzes the condensation of N-acetylmannosamine and phosphoenolpyruvate to form N-acetylneuraminic acid. This report demonstrates the first structural information on sialic acid synthase by CD MALDI-TOF and chemical cross-linking studies. Also a specific cleavage by endogenous protease(s) has been identified at Lys280 of the enzyme (40 kDa) by LC-MS and N-terminal sequencing analyses. The cleavage results in the formation of two inactive fragments of 33 and 7 kDa. The structural analysis indicates that the fragmentation is associated with a significant change of the enzyme from a tetrameric to trimeric form and alterations in both secondary and native quaternary structures.

AB - Sialic acid synthase encoded by the neuB gene of Escherichia coli catalyzes the condensation of N-acetylmannosamine and phosphoenolpyruvate to form N-acetylneuraminic acid. This report demonstrates the first structural information on sialic acid synthase by CD MALDI-TOF and chemical cross-linking studies. Also a specific cleavage by endogenous protease(s) has been identified at Lys280 of the enzyme (40 kDa) by LC-MS and N-terminal sequencing analyses. The cleavage results in the formation of two inactive fragments of 33 and 7 kDa. The structural analysis indicates that the fragmentation is associated with a significant change of the enzyme from a tetrameric to trimeric form and alterations in both secondary and native quaternary structures.

KW - Circular dichroism (CD)

KW - Matrix-assisted laser desorption (MALDI)/ionization-time-of-flight (TOF)

KW - N-Acetylneuraminic acid (NeuAc)

KW - Sialic acid synthase

KW - Subunit cross-linking

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EP - 173

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 1

ER -

Structural characterization of Escherichia coli sialic acid synthase

Abstract

Keywords

ASJC Scopus subject areas

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