1jm0
From Proteopedia
(New page: 200px<br /><applet load="1jm0" size="450" color="white" frame="true" align="right" spinBox="true" caption="1jm0, resolution 1.70Å" /> '''CRYSTAL STRUCTURE OF...) |
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| - | [[Image:1jm0.jpg|left|200px]]<br /><applet load="1jm0" size=" | + | [[Image:1jm0.jpg|left|200px]]<br /><applet load="1jm0" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1jm0, resolution 1.70Å" /> | caption="1jm0, resolution 1.70Å" /> | ||
'''CRYSTAL STRUCTURE OF FOUR-HELIX BUNDLE MODEL'''<br /> | '''CRYSTAL STRUCTURE OF FOUR-HELIX BUNDLE MODEL'''<br /> | ||
==Overview== | ==Overview== | ||
| - | De novo design of proteins provides an attractive approach to uncover the | + | De novo design of proteins provides an attractive approach to uncover the essential features required for their functions. Previously, we described the design and crystal structure determination of a di-Zn(II) complex of "due-ferri-1" (DF1), a protein patterned after the diiron-dimanganese class of redox-active proteins [Lombardi, A.; Summa, C.; Geremia, S.; Randaccio, L.; Pavone, V.; DeGrado, W. F. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 6298-6305]. The overall structure of DF1, which contains a carboxylate-bridged dinuclear metal site, agrees well with the intended design. However, access to this dimetal site is blocked by a pair of hydrophobic leucine residues (L13 and L13'), which prevent facile entry of metal ions and small molecules. We have now taken the next step in the eventual construction of a catalytically active metalloenzyme by engineering an active site cavity into DF1 through the replacement of these two leucine residues with smaller residues. The crystal structure of the dimanganous form of L13A-DF1 indeed shows a substrate access channel to the dimetal center. In the crystal structure, water molecules and a ligating dimethyl sulfoxide molecule, which forms a monatomic bridge between the metal ions, occupy the cavity. Furthermore, the diferric form of a derivative of L13A-DF1, DF2, is shown to bind azide, acetate, and small aromatic molecules. |
==About this Structure== | ==About this Structure== | ||
| - | 1JM0 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/ ] with MN, ACE, NH2 and DMS as [http://en.wikipedia.org/wiki/ligands ligands]. This structure | + | 1JM0 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/ ] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=ACE:'>ACE</scene>, <scene name='pdbligand=NH2:'>NH2</scene> and <scene name='pdbligand=DMS:'>DMS</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. This structure supersedes the now removed PDB entry 1HR5. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1JM0 OCA]. |
==Reference== | ==Reference== | ||
Toward the de novo design of a catalytically active helix bundle: a substrate-accessible carboxylate-bridged dinuclear metal center., Di Costanzo L, Wade H, Geremia S, Randaccio L, Pavone V, DeGrado WF, Lombardi A, J Am Chem Soc. 2001 Dec 26;123(51):12749-57. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11749531 11749531] | Toward the de novo design of a catalytically active helix bundle: a substrate-accessible carboxylate-bridged dinuclear metal center., Di Costanzo L, Wade H, Geremia S, Randaccio L, Pavone V, DeGrado WF, Lombardi A, J Am Chem Soc. 2001 Dec 26;123(51):12749-57. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=11749531 11749531] | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
| - | [[Category: Costanzo, L | + | [[Category: Costanzo, L Di.]] |
[[Category: Geremia, S.]] | [[Category: Geremia, S.]] | ||
[[Category: ACE]] | [[Category: ACE]] | ||
| Line 21: | Line 21: | ||
[[Category: protein design]] | [[Category: protein design]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:24:10 2008'' |
Revision as of 11:24, 21 February 2008
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CRYSTAL STRUCTURE OF FOUR-HELIX BUNDLE MODEL
Overview
De novo design of proteins provides an attractive approach to uncover the essential features required for their functions. Previously, we described the design and crystal structure determination of a di-Zn(II) complex of "due-ferri-1" (DF1), a protein patterned after the diiron-dimanganese class of redox-active proteins [Lombardi, A.; Summa, C.; Geremia, S.; Randaccio, L.; Pavone, V.; DeGrado, W. F. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 6298-6305]. The overall structure of DF1, which contains a carboxylate-bridged dinuclear metal site, agrees well with the intended design. However, access to this dimetal site is blocked by a pair of hydrophobic leucine residues (L13 and L13'), which prevent facile entry of metal ions and small molecules. We have now taken the next step in the eventual construction of a catalytically active metalloenzyme by engineering an active site cavity into DF1 through the replacement of these two leucine residues with smaller residues. The crystal structure of the dimanganous form of L13A-DF1 indeed shows a substrate access channel to the dimetal center. In the crystal structure, water molecules and a ligating dimethyl sulfoxide molecule, which forms a monatomic bridge between the metal ions, occupy the cavity. Furthermore, the diferric form of a derivative of L13A-DF1, DF2, is shown to bind azide, acetate, and small aromatic molecules.
About this Structure
1JM0 is a Protein complex structure of sequences from [1] with , , and as ligands. This structure supersedes the now removed PDB entry 1HR5. Full crystallographic information is available from OCA.
Reference
Toward the de novo design of a catalytically active helix bundle: a substrate-accessible carboxylate-bridged dinuclear metal center., Di Costanzo L, Wade H, Geremia S, Randaccio L, Pavone V, DeGrado WF, Lombardi A, J Am Chem Soc. 2001 Dec 26;123(51):12749-57. PMID:11749531
Page seeded by OCA on Thu Feb 21 13:24:10 2008
Categories: Protein complex | Costanzo, L Di. | Geremia, S. | ACE | DMS | MN | NH2 | Alpha-helical bundle | Protein design
