1bl4
From Proteopedia
(New page: 200px<br /><applet load="1bl4" size="450" color="white" frame="true" align="right" spinBox="true" caption="1bl4, resolution 1.9Å" /> '''FKBP MUTANT F36V COMP...) |
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| - | [[Image:1bl4.gif|left|200px]]<br /><applet load="1bl4" size=" | + | [[Image:1bl4.gif|left|200px]]<br /><applet load="1bl4" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1bl4, resolution 1.9Å" /> | caption="1bl4, resolution 1.9Å" /> | ||
'''FKBP MUTANT F36V COMPLEXED WITH REMODELED SYNTHETIC LIGAND'''<br /> | '''FKBP MUTANT F36V COMPLEXED WITH REMODELED SYNTHETIC LIGAND'''<br /> | ||
==Overview== | ==Overview== | ||
| - | FKBP ligand homodimers can be used to activate signaling events inside | + | FKBP ligand homodimers can be used to activate signaling events inside cells and animals that have been engineered to express fusions between appropriate signaling domains and FKBP. However, use of these dimerizers in vivo is potentially limited by ligand binding to endogenous FKBP. We have designed ligands that bind specifically to a mutated FKBP over the wild-type protein by remodeling an FKBP-ligand interface to introduce a specificity binding pocket. A compound bearing an ethyl substituent in place of a carbonyl group exhibited sub-nanomolar affinity and 1,000-fold selectivity for a mutant FKBP with a compensating truncation of a phenylalanine residue. Structural and functional analysis of the new pocket showed that recognition is surprisingly relaxed, with the modified ligand only partially filling the engineered cavity. We incorporated the specificity pocket into a fusion protein containing FKBP and the intracellular domain of the Fas receptor. Cells expressing this modified chimeric protein potently underwent apoptosis in response to AP1903, a homodimer of the modified ligand, both in culture and when implanted into mice. Remodeled dimerizers such as AP1903 are ideal reagents for controlling the activities of cells that have been modified by gene therapy procedures, without interference from endogenous FKBP. |
==About this Structure== | ==About this Structure== | ||
| - | 1BL4 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with AP1 as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Peptidylprolyl_isomerase Peptidylprolyl isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.2.1.8 5.2.1.8] Full crystallographic information is available from [http:// | + | 1BL4 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=AP1:'>AP1</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Peptidylprolyl_isomerase Peptidylprolyl isomerase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.2.1.8 5.2.1.8] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1BL4 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Amara, J.]] | [[Category: Amara, J.]] | ||
[[Category: Clackson, T.]] | [[Category: Clackson, T.]] | ||
| - | [[Category: Courage, N | + | [[Category: Courage, N L.]] |
[[Category: Gilman, M.]] | [[Category: Gilman, M.]] | ||
| - | [[Category: Hatada, M | + | [[Category: Hatada, M H.]] |
[[Category: Holt, D.]] | [[Category: Holt, D.]] | ||
| - | [[Category: Junior, F | + | [[Category: Junior, F Cerasoli.]] |
[[Category: Lu, X.]] | [[Category: Lu, X.]] | ||
| - | [[Category: Magari, S | + | [[Category: Magari, S R.]] |
| - | [[Category: Rollins, C | + | [[Category: Rollins, C T.]] |
| - | [[Category: Rozamus, L | + | [[Category: Rozamus, L W.]] |
| - | [[Category: Stevenson, L | + | [[Category: Stevenson, L F.]] |
| - | [[Category: Wood, S | + | [[Category: Wood, S A.]] |
[[Category: Yang, W.]] | [[Category: Yang, W.]] | ||
[[Category: AP1]] | [[Category: AP1]] | ||
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[[Category: rotamase]] | [[Category: rotamase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:56:29 2008'' |
Revision as of 09:56, 21 February 2008
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FKBP MUTANT F36V COMPLEXED WITH REMODELED SYNTHETIC LIGAND
Overview
FKBP ligand homodimers can be used to activate signaling events inside cells and animals that have been engineered to express fusions between appropriate signaling domains and FKBP. However, use of these dimerizers in vivo is potentially limited by ligand binding to endogenous FKBP. We have designed ligands that bind specifically to a mutated FKBP over the wild-type protein by remodeling an FKBP-ligand interface to introduce a specificity binding pocket. A compound bearing an ethyl substituent in place of a carbonyl group exhibited sub-nanomolar affinity and 1,000-fold selectivity for a mutant FKBP with a compensating truncation of a phenylalanine residue. Structural and functional analysis of the new pocket showed that recognition is surprisingly relaxed, with the modified ligand only partially filling the engineered cavity. We incorporated the specificity pocket into a fusion protein containing FKBP and the intracellular domain of the Fas receptor. Cells expressing this modified chimeric protein potently underwent apoptosis in response to AP1903, a homodimer of the modified ligand, both in culture and when implanted into mice. Remodeled dimerizers such as AP1903 are ideal reagents for controlling the activities of cells that have been modified by gene therapy procedures, without interference from endogenous FKBP.
About this Structure
1BL4 is a Single protein structure of sequence from Homo sapiens with as ligand. Active as Peptidylprolyl isomerase, with EC number 5.2.1.8 Full crystallographic information is available from OCA.
Reference
Redesigning an FKBP-ligand interface to generate chemical dimerizers with novel specificity., Clackson T, Yang W, Rozamus LW, Hatada M, Amara JF, Rollins CT, Stevenson LF, Magari SR, Wood SA, Courage NL, Lu X, Cerasoli F Jr, Gilman M, Holt DA, Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10437-42. PMID:9724721
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