Expression, purification, characterization and crystallization of flap endonuclease-1 from Methanococcus jannaschii.
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 P2(1) with unit cell dimensions of a = 58.93 A, b = 42.53 A, c = 62.62 A and beta = 92.250. A complete data set has been collected at 2.0 A resolution using a frozen crystal.
Bae
K W
KW
Structural Biology Center, Korea Institute of Science & Technology, Seoul.
Baek
K W
KW
Cho
C S
CS
Hwang
K Y
KY
Kim
H R
HR
Sung
H C
HC
Cho
Y
Y
D015153:Blotting, Western; D003460:Crystallization; D018360:Crystallography, X-Ray; D004260:DNA Repair; D004269:DNA, Bacterial / Q000032:analysis; D004706:Endodeoxyribonucleases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D045585:Flap Endonucleases; D017017:Methanococcus / Q000737:chemistry* / Q000235:genetics
D004269:DNA, Bacterial; D004706:Endodeoxyribonucleases; D045585:Flap Endonucleases