Cloning and functional expression of a novel GDP-6-deoxy-D-talose synthetase from Actinobacillus actinomycetemcomitans

Minna Mäki (Corresponding Author), Nina Järvinen, Jarkko Räbinä, Hannu Maaheimo, Pirkko Mattila, Risto Renkonen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

15 Citations (Scopus)

Abstract

Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus that can cause various forms of severe periodontitis and other nonoral infections in human patients. The serotype a–specific polysaccharide antigen of A. actinomycetemcomitans contains solely 6-deoxy-D-talose and its O-2 acetylated modification. This polysaccharide is synthesized from the donor GDP-6-deoxy-D-talose with the relevant talosylation enzyme(s). In the synthesis of GDP-6- deoxy-D-talose, GDP-D-mannose is first converted by GDP-mannose-4,6-dehydratase (GMD) to GDP-4-keto-6-deoxy-D-mannose and then reduced to GDP-6-deoxy-D-talose by GDP-6-deoxy-D-talose synthetase (GTS). In this study, we cloned and overexpressed in Escherichia coli the A. actinomycetemcomitans GTS enzyme responsible for the synthesis of GDP-6-deoxy-D-talose. The recombinant A. actinomycetemcomitans GTS enzyme expressed in E. coli converted the GDP-4-keto-6-deoxy-intermediate to a novel GDP-deoxyhexose. The synthesized GDP-deoxyhexose was shown to be GDP-6-deoxy-D-talose by HPLC, MALDI-TOF MS, and NMR spectroscopy. The functional expression of gts provides another enzymatically defined pathway for the synthesis of GDP-deoxyhexoses, which can be used as donors for the corresponding glycosyltransferases
Original languageEnglish
Pages (from-to)295-303
Number of pages9
JournalGlycobiology
Volume13
Issue number4
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Cloning
Organism Cloning
Guanosine Diphosphate Mannose
Aggregatibacter actinomycetemcomitans
Mannose
Escherichia coli
Polysaccharides
Tissue Donors
Hydro-Lyases
Rhamnose
Glycosyltransferases
Periodontitis
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Ligases
Actinobacillus actinomycetemcomitans GDP-6-deoxy-D-talose synthetase
L-talo-gamma-lactone
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
High Pressure Liquid Chromatography
Antigens

Keywords

  • Actinobacillus actinomycetemcomitans
  • coccobacillus
  • periodontitis
  • synthetase
  • talose
  • enzymes

Cite this

Mäki, Minna ; Järvinen, Nina ; Räbinä, Jarkko ; Maaheimo, Hannu ; Mattila, Pirkko ; Renkonen, Risto. / Cloning and functional expression of a novel GDP-6-deoxy-D-talose synthetase from Actinobacillus actinomycetemcomitans. In: Glycobiology. 2003 ; Vol. 13, No. 4. pp. 295-303.
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Cloning and functional expression of a novel GDP-6-deoxy-D-talose synthetase from Actinobacillus actinomycetemcomitans. / Mäki, Minna (Corresponding Author); Järvinen, Nina; Räbinä, Jarkko; Maaheimo, Hannu; Mattila, Pirkko; Renkonen, Risto (Corresponding Author).

In: Glycobiology, Vol. 13, No. 4, 2003, p. 295-303.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cloning and functional expression of a novel GDP-6-deoxy-D-talose synthetase from Actinobacillus actinomycetemcomitans

AU - Mäki, Minna

AU - Järvinen, Nina

AU - Räbinä, Jarkko

AU - Maaheimo, Hannu

AU - Mattila, Pirkko

AU - Renkonen, Risto

PY - 2003

Y1 - 2003

N2 - Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus that can cause various forms of severe periodontitis and other nonoral infections in human patients. The serotype a–specific polysaccharide antigen of A. actinomycetemcomitans contains solely 6-deoxy-D-talose and its O-2 acetylated modification. This polysaccharide is synthesized from the donor GDP-6-deoxy-D-talose with the relevant talosylation enzyme(s). In the synthesis of GDP-6- deoxy-D-talose, GDP-D-mannose is first converted by GDP-mannose-4,6-dehydratase (GMD) to GDP-4-keto-6-deoxy-D-mannose and then reduced to GDP-6-deoxy-D-talose by GDP-6-deoxy-D-talose synthetase (GTS). In this study, we cloned and overexpressed in Escherichia coli the A. actinomycetemcomitans GTS enzyme responsible for the synthesis of GDP-6-deoxy-D-talose. The recombinant A. actinomycetemcomitans GTS enzyme expressed in E. coli converted the GDP-4-keto-6-deoxy-intermediate to a novel GDP-deoxyhexose. The synthesized GDP-deoxyhexose was shown to be GDP-6-deoxy-D-talose by HPLC, MALDI-TOF MS, and NMR spectroscopy. The functional expression of gts provides another enzymatically defined pathway for the synthesis of GDP-deoxyhexoses, which can be used as donors for the corresponding glycosyltransferases

AB - Actinobacillus actinomycetemcomitans is a Gram-negative coccobacillus that can cause various forms of severe periodontitis and other nonoral infections in human patients. The serotype a–specific polysaccharide antigen of A. actinomycetemcomitans contains solely 6-deoxy-D-talose and its O-2 acetylated modification. This polysaccharide is synthesized from the donor GDP-6-deoxy-D-talose with the relevant talosylation enzyme(s). In the synthesis of GDP-6- deoxy-D-talose, GDP-D-mannose is first converted by GDP-mannose-4,6-dehydratase (GMD) to GDP-4-keto-6-deoxy-D-mannose and then reduced to GDP-6-deoxy-D-talose by GDP-6-deoxy-D-talose synthetase (GTS). In this study, we cloned and overexpressed in Escherichia coli the A. actinomycetemcomitans GTS enzyme responsible for the synthesis of GDP-6-deoxy-D-talose. The recombinant A. actinomycetemcomitans GTS enzyme expressed in E. coli converted the GDP-4-keto-6-deoxy-intermediate to a novel GDP-deoxyhexose. The synthesized GDP-deoxyhexose was shown to be GDP-6-deoxy-D-talose by HPLC, MALDI-TOF MS, and NMR spectroscopy. The functional expression of gts provides another enzymatically defined pathway for the synthesis of GDP-deoxyhexoses, which can be used as donors for the corresponding glycosyltransferases

KW - Actinobacillus actinomycetemcomitans

KW - coccobacillus

KW - periodontitis

KW - synthetase

KW - talose

KW - enzymes

U2 - 10.1093/glycob/cwg035

DO - 10.1093/glycob/cwg035

M3 - Article

VL - 13

SP - 295

EP - 303

JO - Glycobiology

JF - Glycobiology

SN - 0959-6658

IS - 4

ER -