2vh5: Difference between revisions

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==Overview==

~~Many disease-related processes occur via protein complexes~~ that are~~, considered undruggable with small molecules. An example is RAS, which is, frequently mutated~~ in ~~cancer~~ and ~~contributes to initiation and maintenance~~, of the ~~disease by constitutive signal transduction through protein, interaction with effector proteins, like PI3K, RAF and RALGDS. Such, protein interactions are therefore significant targets for therapy~~. We~~, describe~~ a ~~single immunoglobulin variable region domain that specifically, binds to activated GTP-bound RAS and prevents RAS~~-~~dependent tumorigenesis, in a mouse model. The~~ crystal structure of the ~~immunoglobulin~~-~~RAS complex, shows that~~ the variable ~~region competitively binds to~~ the ~~conformationally, variant regions~~ of ~~RAS~~, ~~where its signalling effector molecules interact.~~, ~~This allows~~ the ~~plasma membrane targeted single~~ domain ~~intrabody~~ to~~, inhibit signalling by mutant RAS. This mode~~ of ~~action is a novel advance,~~ to ~~directly interfere~~ with ~~oncogenic RAS~~ function ~~in human cancer and, shows a universally applicable approach to develop macromolecules~~ to, ~~combat cancer. In addition, this method illustrates~~ a ~~general means~~ for~~, interfering with protein interactions that are commonly considered, intractable as conventional drug targets~~.

Intracellular antibody fragments that interfere with molecular interactions inside cells are valuable in investigation of interactomes and in therapeutics, but their application demands that they function in the reducing cellular milieu. We show here a 2.7-A crystal structure of intracellular antibody folds based on scaffolds developed from intracellular antibody capture technology, and we reveal that there is no structural or functional difference with or without the intra-domain disulfide bond of the variable domain of heavy chain or the variable domain of light chain. The data indicate that, in the reducing in vivo environment, the absence of the intra-domain disulfide bond is not an impediment to correction of antibody folding or to interaction with antigen. Thus, the structural constraints for in-cell function are intrinsic to variable single-domain framework sequences, providing a generic scaffold for isolation of functional intracellular antibody single domains.

==About this Structure==

2VH5 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=ZN:'>ZN</scene>, <scene name='pdbligand=MG:'>MG</scene> and <scene name='pdbligand=GTP:'>GTP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Known structural/functional Sites: <scene name='pdbsite=AC1:Gtp Binding Site For Chain R'>AC1</scene>, <scene name='pdbsite=AC2:Mg Binding Site For Chain R'>AC2</scene> and <scene name='pdbsite=AC3:Zn Binding Site For Chain H'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VH5 OCA].

2VH5 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=ZN:'>ZN</scene>, <scene name='pdbligand=MG:'>MG</scene> and <scene name='pdbligand=GTP:'>GTP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Known structural/functional Sites: <scene name='pdbsite=AC1:Gtp+Binding+Site+For+Chain+R'>AC1</scene>, <scene name='pdbsite=AC2:Mg+Binding+Site+For+Chain+R'>AC2</scene> and <scene name='pdbsite=AC3:Zn+Binding+Site+For+Chain+H'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VH5 OCA].

==Reference==

~~Tumour prevention by a single~~ antibody domain ~~targeting the interaction of signal transduction proteins with RAS~~., Tanaka T~~, Williams RL~~, Rabbitts TH, ~~EMBO~~ J. ~~2007 Jul 11~~;26(13):~~3250~~-9. Epub 2007 ~~Jun 14~~. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=~~17568777 17568777~~]

Functional intracellular antibody fragments do not require invariant intra-domain disulfide bonds., Tanaka T, Rabbitts TH, J Mol Biol. 2008 Feb 22;376(3):749-57. Epub 2007 Dec 4. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18187153 18187153]

[[Category: Homo sapiens]]

[[Category: Protein complex]]

[[Category: Rabbitts, T.H.]]

[[Category: Rabbitts, T H.]]

[[Category: Tanaka, T.]]

[[Category: Williams, R.L.]]

[[Category: Williams, R L.]]

[[Category: GTP]]

[[Category: MG]]

Line 39:

[[Category: signaling protein/immune system]]

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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:56:10 2008''

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@@ Line 4: / Line 4: @@
 ==Overview==
-Many disease-related processes occur via protein complexes that are, considered undruggable with small molecules. An example is RAS, which is, frequently mutated in cancer and contributes to initiation and maintenance, of the disease by constitutive signal transduction through protein, interaction with effector proteins, like PI3K, RAF and RALGDS. Such, protein interactions are therefore significant targets for therapy. We, describe a single immunoglobulin variable region domain that specifically, binds to activated GTP-bound RAS and prevents RAS-dependent tumorigenesis, in a mouse model. The crystal structure of the immunoglobulin-RAS complex, shows that the variable region competitively binds to the conformationally, variant regions of RAS, where its signalling effector molecules interact., This allows the plasma membrane targeted single domain intrabody to, inhibit signalling by mutant RAS. This mode of action is a novel advance, to directly interfere with oncogenic RAS function in human cancer and, shows a universally applicable approach to develop macromolecules to, combat cancer. In addition, this method illustrates a general means for, interfering with protein interactions that are commonly considered, intractable as conventional drug targets.
+Intracellular antibody fragments that interfere with molecular interactions inside cells are valuable in investigation of interactomes and in therapeutics, but their application demands that they function in the reducing cellular milieu. We show here a 2.7-A crystal structure of intracellular antibody folds based on scaffolds developed from intracellular antibody capture technology, and we reveal that there is no structural or functional difference with or without the intra-domain disulfide bond of the variable domain of heavy chain or the variable domain of light chain. The data indicate that, in the reducing in vivo environment, the absence of the intra-domain disulfide bond is not an impediment to correction of antibody folding or to interaction with antigen. Thus, the structural constraints for in-cell function are intrinsic to variable single-domain framework sequences, providing a generic scaffold for isolation of functional intracellular antibody single domains.
 ==About this Structure==
-VH5 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=ZN:'>ZN</scene>, <scene name='pdbligand=MG:'>MG</scene> and <scene name='pdbligand=GTP:'>GTP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Known structural/functional Sites: <scene name='pdbsite=AC1:Gtp Binding Site For Chain R'>AC1</scene>, <scene name='pdbsite=AC2:Mg Binding Site For Chain R'>AC2</scene> and <scene name='pdbsite=AC3:Zn Binding Site For Chain H'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VH5 OCA].
+VH5 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=ZN:'>ZN</scene>, <scene name='pdbligand=MG:'>MG</scene> and <scene name='pdbligand=GTP:'>GTP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Known structural/functional Sites: <scene name='pdbsite=AC1:Gtp+Binding+Site+For+Chain+R'>AC1</scene>, <scene name='pdbsite=AC2:Mg+Binding+Site+For+Chain+R'>AC2</scene> and <scene name='pdbsite=AC3:Zn+Binding+Site+For+Chain+H'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VH5 OCA].
 ==Reference==
-Tumour prevention by a single antibody domain targeting the interaction of signal transduction proteins with RAS., Tanaka T, Williams RL, Rabbitts TH, EMBO J. 2007 Jul 11;26(13):3250-9. Epub 2007 Jun 14. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17568777 17568777]
+Functional intracellular antibody fragments do not require invariant intra-domain disulfide bonds., Tanaka T, Rabbitts TH, J Mol Biol. 2008 Feb 22;376(3):749-57. Epub 2007 Dec 4. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18187153 18187153]
 [[Category: Homo sapiens]]
 [[Category: Protein complex]]
-[[Category: Rabbitts, T.H.]]
+[[Category: Rabbitts, T H.]]
 [[Category: Tanaka, T.]]
-[[Category: Williams, R.L.]]
+[[Category: Williams, R L.]]
 [[Category: GTP]]
 [[Category: MG]]
@@ Line 39: / Line 39: @@
 [[Category: signaling protein/immune system]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 10:56:59 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:56:10 2008''