1q1s: Difference between revisions
New page: left|200px<br /><applet load="1q1s" size="450" color="white" frame="true" align="right" spinBox="true" caption="1q1s, resolution 2.30Å" /> '''Mouse Importin alpha... |
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[[Image:1q1s.gif|left|200px]]<br /><applet load="1q1s" size=" | [[Image:1q1s.gif|left|200px]]<br /><applet load="1q1s" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1q1s, resolution 2.30Å" /> | caption="1q1s, resolution 2.30Å" /> | ||
'''Mouse Importin alpha- phosphorylated SV40 CN peptide complex'''<br /> | '''Mouse Importin alpha- phosphorylated SV40 CN peptide complex'''<br /> | ||
==Overview== | ==Overview== | ||
The nuclear import of simian-virus-40 large T-antigen (tumour antigen) is | The nuclear import of simian-virus-40 large T-antigen (tumour antigen) is enhanced via phosphorylation by the protein kinase CK2 at Ser112 in the vicinity of the NLS (nuclear localization sequence). To determine the structural basis of the effect of the sequences flanking the basic cluster KKKRK, and the effect of phosphorylation on the recognition of the NLS by the nuclear import factor importin-alpha (Impalpha), we co-crystallized non-autoinhibited Impalpha with peptides corresponding to the phosphorylated and non-phosphorylated forms of the NLS, and determined the crystal structures of the complexes. The structures show that the amino acids N-terminally flanking the basic cluster make specific contacts with the receptor that are distinct from the interactions between bipartite NLSs and Impalpha. We confirm the important role of flanking sequences using binding assays. Unexpectedly, the regions of the peptides containing the phosphorylation site do not make specific contacts with the receptor. Binding assays confirm that phosphorylation does not increase the affinity of the T-antigen NLS to Impalpha. We conclude that the sequences flanking the basic clusters in NLSs play a crucial role in nuclear import by modulating the recognition of the NLS by Impalpha, whereas phosphorylation of the T-antigen enhances nuclear import by a mechanism that does not involve a direct interaction of the phosphorylated residue with Impalpha. | ||
==About this Structure== | ==About this Structure== | ||
1Q1S is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http:// | 1Q1S is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Q1S OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Mus musculus]] | [[Category: Mus musculus]] | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
[[Category: Fontes, M | [[Category: Fontes, M R.M.]] | ||
[[Category: Jans, D | [[Category: Jans, D A.]] | ||
[[Category: John, A.]] | [[Category: John, A.]] | ||
[[Category: Kobe, B.]] | [[Category: Kobe, B.]] | ||
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[[Category: x-ray crystal structure]] | [[Category: x-ray crystal structure]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:34:57 2008'' | ||
Revision as of 15:35, 21 February 2008
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Mouse Importin alpha- phosphorylated SV40 CN peptide complex
OverviewOverview
The nuclear import of simian-virus-40 large T-antigen (tumour antigen) is enhanced via phosphorylation by the protein kinase CK2 at Ser112 in the vicinity of the NLS (nuclear localization sequence). To determine the structural basis of the effect of the sequences flanking the basic cluster KKKRK, and the effect of phosphorylation on the recognition of the NLS by the nuclear import factor importin-alpha (Impalpha), we co-crystallized non-autoinhibited Impalpha with peptides corresponding to the phosphorylated and non-phosphorylated forms of the NLS, and determined the crystal structures of the complexes. The structures show that the amino acids N-terminally flanking the basic cluster make specific contacts with the receptor that are distinct from the interactions between bipartite NLSs and Impalpha. We confirm the important role of flanking sequences using binding assays. Unexpectedly, the regions of the peptides containing the phosphorylation site do not make specific contacts with the receptor. Binding assays confirm that phosphorylation does not increase the affinity of the T-antigen NLS to Impalpha. We conclude that the sequences flanking the basic clusters in NLSs play a crucial role in nuclear import by modulating the recognition of the NLS by Impalpha, whereas phosphorylation of the T-antigen enhances nuclear import by a mechanism that does not involve a direct interaction of the phosphorylated residue with Impalpha.
About this StructureAbout this Structure
1Q1S is a Protein complex structure of sequences from Mus musculus. Full crystallographic information is available from OCA.
ReferenceReference
Role of flanking sequences and phosphorylation in the recognition of the simian-virus-40 large T-antigen nuclear localization sequences by importin-alpha., Fontes MR, Teh T, Toth G, John A, Pavo I, Jans DA, Kobe B, Biochem J. 2003 Oct 15;375(Pt 2):339-49. PMID:12852786
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