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1ywt

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(New page: 200px<br /> <applet load="1ywt" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ywt, resolution 2.40&Aring;" /> '''Crystal structure o...)
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<applet load="1ywt" size="450" color="white" frame="true" align="right" spinBox="true"
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'''Crystal structure of the human sigma isoform of 14-3-3 in complex with a mode-1 phosphopeptide'''<br />
'''Crystal structure of the human sigma isoform of 14-3-3 in complex with a mode-1 phosphopeptide'''<br />
==Overview==
==Overview==
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The 14-3-3 family of proteins includes seven isotypes in mammalian cells, that play numerous diverse roles in intracellular signaling. Most 14-3-3, proteins form homodimers and mixed heterodimers between different, isotypes, with overlapping roles in ligand binding. In contrast, one, mammalian isoform, 14-3-3sigma, expressed primarily in epithelial cells, appears to play a unique role in the cellular response to DNA damage and, in human oncogenesis. The biological and structural basis for these, 14-3-3sigma-specific functions is unknown. We demonstrate that endogenous, 14-3-3sigma preferentially forms homodimers in cells. We have solved the, x-ray crystal structure of 14-3-3sigma bound to an optimal phosphopeptide, ligand at 2.4 angstroms resolution. The structure reveals the presence of, stabilizing ring-ring and salt bridge interactions unique to the, 14-3-3sigma homodimer structure and potentially destabilizing, electrostatic interactions between subunits in 14-3-3sigma-containing, heterodimers, rationalizing preferential homodimerization of 14-3-3sigma, in vivo. The interaction of the phosphopeptide with 14-3-3 reveals a, conserved mechanism for phospho-dependent ligand binding, implying that, the phosphopeptide binding cleft is not the critical determinant of the, unique biological properties of 14-3-3sigma. Instead, the structure, suggests a second ligand binding site involved in 14-3-3sigma-specific, ligand discrimination. We have confirmed this by site-directed mutagenesis, of three sigma-specific residues that uniquely define this site. Mutation, of these residues to the alternative sequence that is absolutely conserved, in all other 14-3-3 isotypes confers upon 14-3-3sigma the ability to bind, to Cdc25C, a ligand that is known to bind to other 14-3-3 proteins but not, to sigma.
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The 14-3-3 family of proteins includes seven isotypes in mammalian cells that play numerous diverse roles in intracellular signaling. Most 14-3-3 proteins form homodimers and mixed heterodimers between different isotypes, with overlapping roles in ligand binding. In contrast, one mammalian isoform, 14-3-3sigma, expressed primarily in epithelial cells, appears to play a unique role in the cellular response to DNA damage and in human oncogenesis. The biological and structural basis for these 14-3-3sigma-specific functions is unknown. We demonstrate that endogenous 14-3-3sigma preferentially forms homodimers in cells. We have solved the x-ray crystal structure of 14-3-3sigma bound to an optimal phosphopeptide ligand at 2.4 angstroms resolution. The structure reveals the presence of stabilizing ring-ring and salt bridge interactions unique to the 14-3-3sigma homodimer structure and potentially destabilizing electrostatic interactions between subunits in 14-3-3sigma-containing heterodimers, rationalizing preferential homodimerization of 14-3-3sigma in vivo. The interaction of the phosphopeptide with 14-3-3 reveals a conserved mechanism for phospho-dependent ligand binding, implying that the phosphopeptide binding cleft is not the critical determinant of the unique biological properties of 14-3-3sigma. Instead, the structure suggests a second ligand binding site involved in 14-3-3sigma-specific ligand discrimination. We have confirmed this by site-directed mutagenesis of three sigma-specific residues that uniquely define this site. Mutation of these residues to the alternative sequence that is absolutely conserved in all other 14-3-3 isotypes confers upon 14-3-3sigma the ability to bind to Cdc25C, a ligand that is known to bind to other 14-3-3 proteins but not to sigma.
==About this Structure==
==About this Structure==
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1YWT 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 CA as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1YWT OCA].
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1YWT 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=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1YWT OCA].
==Reference==
==Reference==
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[[Category: Homo sapiens]]
[[Category: Homo sapiens]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Artim, S.C.]]
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[[Category: Artim, S C.]]
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[[Category: Grant, R.A.]]
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[[Category: Grant, R A.]]
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[[Category: Wilker, E.W.]]
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[[Category: Wilker, E W.]]
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[[Category: Yaffe, M.B.]]
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[[Category: Yaffe, M B.]]
[[Category: CA]]
[[Category: CA]]
[[Category: 14-3-3]]
[[Category: 14-3-3]]
[[Category: protein-phosphopeptide complex]]
[[Category: protein-phosphopeptide complex]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 20:25:54 2007''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:09:55 2008''

Revision as of 14:09, 21 February 2008


1ywt, resolution 2.40Å

Drag the structure with the mouse to rotate

Crystal structure of the human sigma isoform of 14-3-3 in complex with a mode-1 phosphopeptide

Overview

The 14-3-3 family of proteins includes seven isotypes in mammalian cells that play numerous diverse roles in intracellular signaling. Most 14-3-3 proteins form homodimers and mixed heterodimers between different isotypes, with overlapping roles in ligand binding. In contrast, one mammalian isoform, 14-3-3sigma, expressed primarily in epithelial cells, appears to play a unique role in the cellular response to DNA damage and in human oncogenesis. The biological and structural basis for these 14-3-3sigma-specific functions is unknown. We demonstrate that endogenous 14-3-3sigma preferentially forms homodimers in cells. We have solved the x-ray crystal structure of 14-3-3sigma bound to an optimal phosphopeptide ligand at 2.4 angstroms resolution. The structure reveals the presence of stabilizing ring-ring and salt bridge interactions unique to the 14-3-3sigma homodimer structure and potentially destabilizing electrostatic interactions between subunits in 14-3-3sigma-containing heterodimers, rationalizing preferential homodimerization of 14-3-3sigma in vivo. The interaction of the phosphopeptide with 14-3-3 reveals a conserved mechanism for phospho-dependent ligand binding, implying that the phosphopeptide binding cleft is not the critical determinant of the unique biological properties of 14-3-3sigma. Instead, the structure suggests a second ligand binding site involved in 14-3-3sigma-specific ligand discrimination. We have confirmed this by site-directed mutagenesis of three sigma-specific residues that uniquely define this site. Mutation of these residues to the alternative sequence that is absolutely conserved in all other 14-3-3 isotypes confers upon 14-3-3sigma the ability to bind to Cdc25C, a ligand that is known to bind to other 14-3-3 proteins but not to sigma.

About this Structure

1YWT is a Single protein structure of sequence from Homo sapiens with as ligand. Full crystallographic information is available from OCA.

Reference

A structural basis for 14-3-3sigma functional specificity., Wilker EW, Grant RA, Artim SC, Yaffe MB, J Biol Chem. 2005 May 13;280(19):18891-8. Epub 2005 Feb 24. PMID:15731107

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