1mj0
From Proteopedia
(New page: 200px<br /><applet load="1mj0" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mj0, resolution 2.031Å" /> '''SANK E3_5: an artif...) |
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| - | [[Image:1mj0.gif|left|200px]]<br /><applet load="1mj0" size=" | + | [[Image:1mj0.gif|left|200px]]<br /><applet load="1mj0" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1mj0, resolution 2.031Å" /> | caption="1mj0, resolution 2.031Å" /> | ||
'''SANK E3_5: an artificial Ankyrin repeat protein'''<br /> | '''SANK E3_5: an artificial Ankyrin repeat protein'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Ankyrin repeat (AR) proteins mediate innumerable protein-protein | + | Ankyrin repeat (AR) proteins mediate innumerable protein-protein interactions in virtually all phyla. This finding suggested the use of AR proteins as designed binding molecules. Based on sequence and structural analyses, we designed a consensus AR with fixed framework and randomized interacting residues. We generated several combinatorial libraries of AR proteins consisting of defined numbers of this repeat. Randomly chosen library members are expressed in soluble form in the cytoplasm of Escherichia coli constituting up to 30% of total cellular protein and show high thermodynamic stability. We determined the crystal structure of one of those library members to 2.0-A resolution, providing insight into the consensus AR fold. Besides the highly complementary hydrophobic repeat-repeat interfaces and the absence of structural irregularities in the consensus AR protein, the regular and extended hydrogen bond networks in the beta-turn and loop regions are noteworthy. Furthermore, all residues found in the turn region of the Ramachandran plot are glycines. Many of these features also occur in natural AR proteins, but not in this rigorous and standardized fashion. We conclude that the AR domain fold is an intrinsically very stable and well-expressed scaffold, able to display randomized interacting residues. This scaffold represents an excellent basis for the design of novel binding molecules. |
==About this Structure== | ==About this Structure== | ||
| - | 1MJ0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Designed_synthetic_gene Designed synthetic gene] with SO4 and TRS as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http:// | + | 1MJ0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Designed_synthetic_gene Designed synthetic gene] with <scene name='pdbligand=SO4:'>SO4</scene> and <scene name='pdbligand=TRS:'>TRS</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MJ0 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Designed synthetic gene]] | [[Category: Designed synthetic gene]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Binz, H | + | [[Category: Binz, H K.]] |
[[Category: Forrer, P.]] | [[Category: Forrer, P.]] | ||
| - | [[Category: Gruetter, M | + | [[Category: Gruetter, M G.]] |
[[Category: Kohl, A.]] | [[Category: Kohl, A.]] | ||
[[Category: Plueckthun, A.]] | [[Category: Plueckthun, A.]] | ||
| - | [[Category: Stumpp, M | + | [[Category: Stumpp, M T.]] |
[[Category: SO4]] | [[Category: SO4]] | ||
[[Category: TRS]] | [[Category: TRS]] | ||
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[[Category: protein engineering]] | [[Category: protein engineering]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:55:38 2008'' |
Revision as of 11:55, 21 February 2008
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SANK E3_5: an artificial Ankyrin repeat protein
Overview
Ankyrin repeat (AR) proteins mediate innumerable protein-protein interactions in virtually all phyla. This finding suggested the use of AR proteins as designed binding molecules. Based on sequence and structural analyses, we designed a consensus AR with fixed framework and randomized interacting residues. We generated several combinatorial libraries of AR proteins consisting of defined numbers of this repeat. Randomly chosen library members are expressed in soluble form in the cytoplasm of Escherichia coli constituting up to 30% of total cellular protein and show high thermodynamic stability. We determined the crystal structure of one of those library members to 2.0-A resolution, providing insight into the consensus AR fold. Besides the highly complementary hydrophobic repeat-repeat interfaces and the absence of structural irregularities in the consensus AR protein, the regular and extended hydrogen bond networks in the beta-turn and loop regions are noteworthy. Furthermore, all residues found in the turn region of the Ramachandran plot are glycines. Many of these features also occur in natural AR proteins, but not in this rigorous and standardized fashion. We conclude that the AR domain fold is an intrinsically very stable and well-expressed scaffold, able to display randomized interacting residues. This scaffold represents an excellent basis for the design of novel binding molecules.
About this Structure
1MJ0 is a Single protein structure of sequence from Designed synthetic gene with and as ligands. Full crystallographic information is available from OCA.
Reference
Designed to be stable: crystal structure of a consensus ankyrin repeat protein., Kohl A, Binz HK, Forrer P, Stumpp MT, Pluckthun A, Grutter MG, Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1700-5. Epub 2003 Feb 3. PMID:12566564
Page seeded by OCA on Thu Feb 21 13:55:38 2008
