Expression, Purification, Characterization and Crystallization of Flap Endonuclease-1 from Methanococcus jannaschii

Keun Wook Bae; Kyu Won Baek; Chun Seok Cho; Kwang Yeon Hwang; Hak Ryul Kim; Ha Chin Sung; Yunje Cho

Expression, Purification, Characterization and Crystallization of Flap Endonuclease-1 from Methanococcus jannaschii

Keun Wook Bae, Kyu Won Baek, Chun Seok Cho, Kwang Yeon Hwang, Hak Ryul Kim, Ha Chin Sung, Yunje Cho

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

Abstract

A gene coding for a protein homologous to the flap endonuclease-1 (FEN-1) was cloned from Methanococcus jannaschii, overexpressed, purified and characterized. The gene product from M. jannaschii shows 5′ endo-/exonuclease and 5′ pseudo-Y-endonuclease activities as observed in the FEN-1 in eukaryotes. In addition, Methanococcus jannaschii FEN-1 functions effectively at high concentrations of salt, unlike eukaryotic FEN-1. We have crystallized Methanococcus jannaschii FEN-1 and analyzed its preliminary character. The crystal belongs to the space group of P2₁ with unit cell dimensions of a = 58.93 Å, b = 42.53 Å, c = 62.62 Å and β = 92.25°. A complete data set has been collected at 2.0 Å resolution using a frozen crystal.

Original language	English
Pages (from-to)	45-48
Number of pages	4
Journal	Molecules and cells
Volume	9
Issue number	1
Publication status	Published - 1999 Feb 28

Keywords

Crystallization
Flap Endonuclease-1
Methanococcus janaschii
Structure Specific Nuclease

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Cite this

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title = "Expression, Purification, Characterization and Crystallization of Flap Endonuclease-1 from Methanococcus jannaschii",

abstract = "A gene coding for a protein homologous to the flap endonuclease-1 (FEN-1) was cloned from Methanococcus jannaschii, overexpressed, purified and characterized. The gene product from M. jannaschii shows 5′ endo-/exonuclease and 5′ pseudo-Y-endonuclease activities as observed in the FEN-1 in eukaryotes. In addition, Methanococcus jannaschii FEN-1 functions effectively at high concentrations of salt, unlike eukaryotic FEN-1. We have crystallized Methanococcus jannaschii FEN-1 and analyzed its preliminary character. The crystal belongs to the space group of P21 with unit cell dimensions of a = 58.93 {\AA}, b = 42.53 {\AA}, c = 62.62 {\AA} and β = 92.25°. A complete data set has been collected at 2.0 {\AA} resolution using a frozen crystal.",

keywords = "Crystallization, Flap Endonuclease-1, Methanococcus janaschii, Structure Specific Nuclease",

author = "Bae, {Keun Wook} and Baek, {Kyu Won} and Cho, {Chun Seok} and Hwang, {Kwang Yeon} and Kim, {Hak Ryul} and Sung, {Ha Chin} and Yunje Cho",

year = "1999",

month = feb,

day = "28",

language = "English",

volume = "9",

pages = "45--48",

journal = "Molecules and cells",

issn = "1016-8478",

publisher = "Elsevier B.V.",

number = "1",

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T1 - Expression, Purification, Characterization and Crystallization of Flap Endonuclease-1 from Methanococcus jannaschii

AU - Bae, Keun Wook

AU - Baek, Kyu Won

AU - Cho, Chun Seok

AU - Hwang, Kwang Yeon

AU - Kim, Hak Ryul

AU - Sung, Ha Chin

AU - Cho, Yunje

PY - 1999/2/28

Y1 - 1999/2/28

N2 - A gene coding for a protein homologous to the flap endonuclease-1 (FEN-1) was cloned from Methanococcus jannaschii, overexpressed, purified and characterized. The gene product from M. jannaschii shows 5′ endo-/exonuclease and 5′ pseudo-Y-endonuclease activities as observed in the FEN-1 in eukaryotes. In addition, Methanococcus jannaschii FEN-1 functions effectively at high concentrations of salt, unlike eukaryotic FEN-1. We have crystallized Methanococcus jannaschii FEN-1 and analyzed its preliminary character. The crystal belongs to the space group of P21 with unit cell dimensions of a = 58.93 Å, b = 42.53 Å, c = 62.62 Å and β = 92.25°. A complete data set has been collected at 2.0 Å resolution using a frozen crystal.

AB - A gene coding for a protein homologous to the flap endonuclease-1 (FEN-1) was cloned from Methanococcus jannaschii, overexpressed, purified and characterized. The gene product from M. jannaschii shows 5′ endo-/exonuclease and 5′ pseudo-Y-endonuclease activities as observed in the FEN-1 in eukaryotes. In addition, Methanococcus jannaschii FEN-1 functions effectively at high concentrations of salt, unlike eukaryotic FEN-1. We have crystallized Methanococcus jannaschii FEN-1 and analyzed its preliminary character. The crystal belongs to the space group of P21 with unit cell dimensions of a = 58.93 Å, b = 42.53 Å, c = 62.62 Å and β = 92.25°. A complete data set has been collected at 2.0 Å resolution using a frozen crystal.

KW - Crystallization

KW - Flap Endonuclease-1

KW - Methanococcus janaschii

KW - Structure Specific Nuclease

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M3 - Article

C2 - 10102570

AN - SCOPUS:0033611614

SN - 1016-8478

VL - 9

SP - 45

EP - 48

JO - Molecules and cells

JF - Molecules and cells

IS - 1

ER -

Expression, Purification, Characterization and Crystallization of Flap Endonuclease-1 from Methanococcus jannaschii

Abstract

Keywords

ASJC Scopus subject areas

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