1mc8
From Proteopedia
(New page: 200px<br /><applet load="1mc8" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mc8, resolution 3.10Å" /> '''Crystal Structure of...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:1mc8.jpg|left|200px]]<br /><applet load="1mc8" size=" | + | [[Image:1mc8.jpg|left|200px]]<br /><applet load="1mc8" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1mc8, resolution 3.10Å" /> | caption="1mc8, resolution 3.10Å" /> | ||
'''Crystal Structure of Flap Endonuclease-1 R42E mutant from Pyrococcus horikoshii'''<br /> | '''Crystal Structure of Flap Endonuclease-1 R42E mutant from Pyrococcus horikoshii'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The crystal structure of flap endonuclease-1 from Pyrococcus horikoshii | + | The crystal structure of flap endonuclease-1 from Pyrococcus horikoshii (phFEN-1) was determined to a resolution of 3.1 A. The active cleft of the phFEN-1 molecule is formed with one large loop and four small loops. We examined the function of the conserved residues and positively charged clusters on these loops by kinetic analysis with 45 different mutants. Arg(40) and Arg(42) on small loop 1, a cluster Lys(193)-Lys(195) on small loop 2, and two sites, Arg(94) and Arg(118)-Lys(119), on the large loop were identified as binding sites. Lys(87) on the large loop may play significant roles in catalytic reaction. Furthermore, we successfully elucidated the function of the four DNA binding sites that form productive ES complexes specific for each endo- or exo-type hydrolysis, probably by bending the substrates. For the endo-activity, Arg(94) and Lys(193)-Lys(195) located at the top and bottom of the molecule were key determinants. For the exo-activity, all four sites were needed, but Arg(118)-Lys(119) was dominant. The major binding sites for both the nick substrate and double-stranded DNA might be the same. |
==About this Structure== | ==About this Structure== | ||
| - | 1MC8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pyrococcus_horikoshii Pyrococcus horikoshii]. Full crystallographic information is available from [http:// | + | 1MC8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pyrococcus_horikoshii Pyrococcus horikoshii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MC8 OCA]. |
==Reference== | ==Reference== | ||
| Line 17: | Line 17: | ||
[[Category: Matsui, E.]] | [[Category: Matsui, E.]] | ||
[[Category: Matsui, I.]] | [[Category: Matsui, I.]] | ||
| - | [[Category: Musti, K | + | [[Category: Musti, K V.]] |
[[Category: Yamazaki, K.]] | [[Category: Yamazaki, K.]] | ||
[[Category: flexible loop]] | [[Category: flexible loop]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:53:46 2008'' |
Revision as of 11:53, 21 February 2008
|
Crystal Structure of Flap Endonuclease-1 R42E mutant from Pyrococcus horikoshii
Overview
The crystal structure of flap endonuclease-1 from Pyrococcus horikoshii (phFEN-1) was determined to a resolution of 3.1 A. The active cleft of the phFEN-1 molecule is formed with one large loop and four small loops. We examined the function of the conserved residues and positively charged clusters on these loops by kinetic analysis with 45 different mutants. Arg(40) and Arg(42) on small loop 1, a cluster Lys(193)-Lys(195) on small loop 2, and two sites, Arg(94) and Arg(118)-Lys(119), on the large loop were identified as binding sites. Lys(87) on the large loop may play significant roles in catalytic reaction. Furthermore, we successfully elucidated the function of the four DNA binding sites that form productive ES complexes specific for each endo- or exo-type hydrolysis, probably by bending the substrates. For the endo-activity, Arg(94) and Lys(193)-Lys(195) located at the top and bottom of the molecule were key determinants. For the exo-activity, all four sites were needed, but Arg(118)-Lys(119) was dominant. The major binding sites for both the nick substrate and double-stranded DNA might be the same.
About this Structure
1MC8 is a Single protein structure of sequence from Pyrococcus horikoshii. Full crystallographic information is available from OCA.
Reference
Molecular structure and novel DNA binding sites located in loops of flap endonuclease-1 from Pyrococcus horikoshii., Matsui E, Musti KV, Abe J, Yamasaki K, Matsui I, Harata K, J Biol Chem. 2002 Oct 4;277(40):37840-7. Epub 2002 Jul 29. PMID:12147694
Page seeded by OCA on Thu Feb 21 13:53:46 2008
