Structural and functional analysis of Vitamin K2 synthesis protein MenD

Amit Priyadarshi, Eunice EunKyeong Kim, Kwang Yeon Hwang

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Here we describe in detail the crystal structures of the Vitamin K2 synthesis protein MenD, from Escherichia coli, in complex with thiamine diphosphate (ThDP) and oxoglutarate, and the effects of cofactor and substrate on its structural stability. This is the first reported structure of MenD in complex with oxoglutarate. The residues Gly472 to Phe488 of the active site region are either disordered, or in an open conformation in the MenD oxoglutarate complex structure, but adopt a closed conformation in the MenD ThDP complex structure. Biospecific-interaction analysis using surface plasmon resonance (SPR) technology reveals an affinity for ThDP and oxoglutarate in the nanomolar range. Biochemical and structural analysis confirmed that MenD is highly dependent on ThDP for its structural stability. Our structural results combined with the biochemical assay reveal novel features of the enzyme that could be utilized in a program of rational structure-based drug design, as well as in helping to enhance our knowledge of the menaquinone synthesis pathway in greater detail.

Original languageEnglish
Pages (from-to)748-751
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume388
Issue number4
DOIs
Publication statusPublished - 2009 Oct 30

Fingerprint

Thiamine Pyrophosphate
Ketoglutaric Acids
Vitamin K 2
Functional analysis
Structural analysis
Conformations
Proteins
Surface Plasmon Resonance
Drug Design
Escherichia coli Proteins
Surface plasmon resonance
Escherichia coli
Assays
Catalytic Domain
Crystal structure
Technology
Substrates
Enzymes
Pharmaceutical Preparations

Keywords

  • Menaquinone
  • MenD
  • Oxoglutarate
  • SPR
  • ThDP
  • Vitamin K

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Structural and functional analysis of Vitamin K2 synthesis protein MenD. / Priyadarshi, Amit; Kim, Eunice EunKyeong; Hwang, Kwang Yeon.

In: Biochemical and Biophysical Research Communications, Vol. 388, No. 4, 30.10.2009, p. 748-751.

Research output: Contribution to journalArticle

@article{5df0ebb6768146a28869b1fd6790421c,
title = "Structural and functional analysis of Vitamin K2 synthesis protein MenD",
abstract = "Here we describe in detail the crystal structures of the Vitamin K2 synthesis protein MenD, from Escherichia coli, in complex with thiamine diphosphate (ThDP) and oxoglutarate, and the effects of cofactor and substrate on its structural stability. This is the first reported structure of MenD in complex with oxoglutarate. The residues Gly472 to Phe488 of the active site region are either disordered, or in an open conformation in the MenD oxoglutarate complex structure, but adopt a closed conformation in the MenD ThDP complex structure. Biospecific-interaction analysis using surface plasmon resonance (SPR) technology reveals an affinity for ThDP and oxoglutarate in the nanomolar range. Biochemical and structural analysis confirmed that MenD is highly dependent on ThDP for its structural stability. Our structural results combined with the biochemical assay reveal novel features of the enzyme that could be utilized in a program of rational structure-based drug design, as well as in helping to enhance our knowledge of the menaquinone synthesis pathway in greater detail.",
keywords = "Menaquinone, MenD, Oxoglutarate, SPR, ThDP, Vitamin K",
author = "Amit Priyadarshi and Kim, {Eunice EunKyeong} and Hwang, {Kwang Yeon}",
year = "2009",
month = "10",
day = "30",
doi = "10.1016/j.bbrc.2009.08.093",
language = "English",
volume = "388",
pages = "748--751",
journal = "The BMJ",
issn = "0730-6512",
publisher = "Kluwer Academic Publishers",
number = "4",

}

TY - JOUR

T1 - Structural and functional analysis of Vitamin K2 synthesis protein MenD

AU - Priyadarshi, Amit

AU - Kim, Eunice EunKyeong

AU - Hwang, Kwang Yeon

PY - 2009/10/30

Y1 - 2009/10/30

N2 - Here we describe in detail the crystal structures of the Vitamin K2 synthesis protein MenD, from Escherichia coli, in complex with thiamine diphosphate (ThDP) and oxoglutarate, and the effects of cofactor and substrate on its structural stability. This is the first reported structure of MenD in complex with oxoglutarate. The residues Gly472 to Phe488 of the active site region are either disordered, or in an open conformation in the MenD oxoglutarate complex structure, but adopt a closed conformation in the MenD ThDP complex structure. Biospecific-interaction analysis using surface plasmon resonance (SPR) technology reveals an affinity for ThDP and oxoglutarate in the nanomolar range. Biochemical and structural analysis confirmed that MenD is highly dependent on ThDP for its structural stability. Our structural results combined with the biochemical assay reveal novel features of the enzyme that could be utilized in a program of rational structure-based drug design, as well as in helping to enhance our knowledge of the menaquinone synthesis pathway in greater detail.

AB - Here we describe in detail the crystal structures of the Vitamin K2 synthesis protein MenD, from Escherichia coli, in complex with thiamine diphosphate (ThDP) and oxoglutarate, and the effects of cofactor and substrate on its structural stability. This is the first reported structure of MenD in complex with oxoglutarate. The residues Gly472 to Phe488 of the active site region are either disordered, or in an open conformation in the MenD oxoglutarate complex structure, but adopt a closed conformation in the MenD ThDP complex structure. Biospecific-interaction analysis using surface plasmon resonance (SPR) technology reveals an affinity for ThDP and oxoglutarate in the nanomolar range. Biochemical and structural analysis confirmed that MenD is highly dependent on ThDP for its structural stability. Our structural results combined with the biochemical assay reveal novel features of the enzyme that could be utilized in a program of rational structure-based drug design, as well as in helping to enhance our knowledge of the menaquinone synthesis pathway in greater detail.

KW - Menaquinone

KW - MenD

KW - Oxoglutarate

KW - SPR

KW - ThDP

KW - Vitamin K

UR - http://www.scopus.com/inward/record.url?scp=69949140290&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69949140290&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2009.08.093

DO - 10.1016/j.bbrc.2009.08.093

M3 - Article

C2 - 19703421

AN - SCOPUS:69949140290

VL - 388

SP - 748

EP - 751

JO - The BMJ

JF - The BMJ

SN - 0730-6512

IS - 4

ER -