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 = "Elsevier BV",

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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U2 - 10.1016/S0006-291X(02)00620-4

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

M3 - Article

C2 - 12083785

AN - SCOPUS:0036305572

SN - 0006-291X

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SP - 167

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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