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Biochem Biophys Res Commun. 2014 Sep 26;452(3):789-94. doi: 10.1016/j.bbrc.2014.08.153. Epub 2014 Sep 6.

Kdo hydroxylase is an inner core assembly enzyme in the Ko-containing lipopolysaccharide biosynthesis.

Author information

1
Center for Theragnosis, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, KIST Campus, Korea University of Science and Technology (UST), Seoul 136-791, Republic of Korea. Electronic address: hschung@kist.re.kr.
2
Center for Theragnosis, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, KIST Campus, Korea University of Science and Technology (UST), Seoul 136-791, Republic of Korea.
3
Department of Biological Chemistry, KIST Campus, Korea University of Science and Technology (UST), Seoul 136-791, Republic of Korea.
4
Center for Theragnosis, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea.
5
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.

Abstract

The lipopolysaccharide (LPS) isolated from certain important Gram-negative pathogens including a human pathogen Yersinia pestis and opportunistic pathogens Burkholderia mallei and Burkholderia pseudomallei contains d-glycero-d-talo-oct-2-ulosonic acid (Ko), an isosteric analog of 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo). Kdo 3-hydroxylase (KdoO), a Fe(2+)/α-KG/O2 dependent dioxygenase from Burkholderia ambifaria and Yersinia pestis is responsible for Ko formation with Kdo2-lipid A as a substrate, but in which stage KdoO functions during the LPS biosynthesis has not been established. Here we purify KdoO from B. ambifaria (BaKdoO) to homogeneity for the first time and characterize its substrates. BaKdoO utilizes Kdo2-lipid IVA or Kdo2-lipid A as a substrate, but not Kdo-lipid IVAin vivo as well as in vitro and Kdo-(Hep)kdo-lipid A in vitro. These data suggest that KdoO is an inner core assembly enzyme that functions after the Kdo-transferase KdtA but before the heptosyl-transferase WaaC enzyme during the Ko-containing LPS biosynthesis.

KEYWORDS:

Burkholderia LPS; Fe(2+)/O(2)/α-KG dependent dioxygenase; Kdo hydroxylase; Ko formation; LPS inner core assembly

PMID:
25204504
PMCID:
PMC4282518
DOI:
10.1016/j.bbrc.2014.08.153
[Indexed for MEDLINE]
Free PMC Article

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