Identification of amino acid residues important for the phosphomannose isomerase activity of PslB in Pseudomonas aeruginosa PAO1

Hui Ju Lee; Hwan You Chang; Nandinin Venkatesan; Hwei Ling Peng

doi:10.1016/j.febslet.2008.09.013

Identification of amino acid residues important for the phosphomannose isomerase activity of PslB in Pseudomonas aeruginosa PAO1

Hui Ju Lee, Hwan You Chang, Nandinin Venkatesan, Hwei Ling Peng^*

^*Corresponding author for this work

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

14 Scopus citations

Abstract

Phosphomannose isomerase (PMI) plays a pivotal role in biosynthesis of GDP-mannose, an important precursor of many polysaccharides. We demonstrate in this study that Pseudomonas aeruginosa pslB encodes a protein with GDP-mannose pyrophosphorylase/PMI dual activities. The PMI activity is Co²⁺-dependent and could be inhibited by GDP-mannose in a competitive manner. Furthermore, the activity could be inactivated by 2,3-butanedione suggesting the presence of a catalytic Arg residue. Site-specific mutations at R373, R472, R479, E410, H411, N433 and E458 increase the K_M approximately 8-20-fold. The PMI activity of PslB was completely diminished with a R408K or R408A, reflecting the importance of this residue in catalysis. Overall, these results provide a basis for understanding the catalytic mechanism of PMI.

Original language	English
Pages (from-to)	3479-3483
Number of pages	5
Journal	FEBS Letters
Volume	582
Issue number	23-24
DOIs	https://doi.org/10.1016/j.febslet.2008.09.013
State	Published - 15 10 2008
Externally published	Yes

Keywords

GDP-mannose pyrophosphorylase
Phosphomannose isomerase
Pseudomonas aeruginosa
Site-directed mutagenesis
pslB

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10.1016/j.febslet.2008.09.013

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@article{7b9419ac213c4be3b7e474453e734c2a,

title = "Identification of amino acid residues important for the phosphomannose isomerase activity of PslB in Pseudomonas aeruginosa PAO1",

abstract = "Phosphomannose isomerase (PMI) plays a pivotal role in biosynthesis of GDP-mannose, an important precursor of many polysaccharides. We demonstrate in this study that Pseudomonas aeruginosa pslB encodes a protein with GDP-mannose pyrophosphorylase/PMI dual activities. The PMI activity is Co2+-dependent and could be inhibited by GDP-mannose in a competitive manner. Furthermore, the activity could be inactivated by 2,3-butanedione suggesting the presence of a catalytic Arg residue. Site-specific mutations at R373, R472, R479, E410, H411, N433 and E458 increase the KM approximately 8-20-fold. The PMI activity of PslB was completely diminished with a R408K or R408A, reflecting the importance of this residue in catalysis. Overall, these results provide a basis for understanding the catalytic mechanism of PMI.",

keywords = "GDP-mannose pyrophosphorylase, Phosphomannose isomerase, Pseudomonas aeruginosa, Site-directed mutagenesis, pslB",

author = "Lee, \{Hui Ju\} and Chang, \{Hwan You\} and Nandinin Venkatesan and Peng, \{Hwei Ling\}",

year = "2008",

month = oct,

day = "15",

doi = "10.1016/j.febslet.2008.09.013",

language = "英语",

volume = "582",

pages = "3479--3483",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc",

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TY - JOUR

T1 - Identification of amino acid residues important for the phosphomannose isomerase activity of PslB in Pseudomonas aeruginosa PAO1

AU - Lee, Hui Ju

AU - Chang, Hwan You

AU - Venkatesan, Nandinin

AU - Peng, Hwei Ling

PY - 2008/10/15

Y1 - 2008/10/15

N2 - Phosphomannose isomerase (PMI) plays a pivotal role in biosynthesis of GDP-mannose, an important precursor of many polysaccharides. We demonstrate in this study that Pseudomonas aeruginosa pslB encodes a protein with GDP-mannose pyrophosphorylase/PMI dual activities. The PMI activity is Co2+-dependent and could be inhibited by GDP-mannose in a competitive manner. Furthermore, the activity could be inactivated by 2,3-butanedione suggesting the presence of a catalytic Arg residue. Site-specific mutations at R373, R472, R479, E410, H411, N433 and E458 increase the KM approximately 8-20-fold. The PMI activity of PslB was completely diminished with a R408K or R408A, reflecting the importance of this residue in catalysis. Overall, these results provide a basis for understanding the catalytic mechanism of PMI.

AB - Phosphomannose isomerase (PMI) plays a pivotal role in biosynthesis of GDP-mannose, an important precursor of many polysaccharides. We demonstrate in this study that Pseudomonas aeruginosa pslB encodes a protein with GDP-mannose pyrophosphorylase/PMI dual activities. The PMI activity is Co2+-dependent and could be inhibited by GDP-mannose in a competitive manner. Furthermore, the activity could be inactivated by 2,3-butanedione suggesting the presence of a catalytic Arg residue. Site-specific mutations at R373, R472, R479, E410, H411, N433 and E458 increase the KM approximately 8-20-fold. The PMI activity of PslB was completely diminished with a R408K or R408A, reflecting the importance of this residue in catalysis. Overall, these results provide a basis for understanding the catalytic mechanism of PMI.

KW - GDP-mannose pyrophosphorylase

KW - Phosphomannose isomerase

KW - Pseudomonas aeruginosa

KW - Site-directed mutagenesis

KW - pslB

UR - https://www.scopus.com/pages/publications/53049083717

U2 - 10.1016/j.febslet.2008.09.013

DO - 10.1016/j.febslet.2008.09.013

M3 - 文章

C2 - 18801364

AN - SCOPUS:53049083717

SN - 0014-5793

VL - 582

SP - 3479

EP - 3483

JO - FEBS Letters

JF - FEBS Letters

IS - 23-24

ER -

Identification of amino acid residues important for the phosphomannose isomerase activity of PslB in Pseudomonas aeruginosa PAO1

Abstract

Keywords

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