Isolation of a point-mutated p47 lacking binding affinity to p97ATPase

Yayoi Kaneko, Kaori Tamura, Go Totsukawa, Hisao Kondo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

p47, a p97-binding protein, functions in Golgi membrane fusion together with p97 and VCIP135, another p97-binding protein. We have succeeded in creating p47 with a point mutation, F253S, which lacks p97-binding affinity. p47 mapping experiments revealed that p47 had two p97-binding regions and the F253S mutation occurred in the first p97-binding site. p47(F253S) could not form a complex with p97 and did not caused any cisternal regrowth in an in vitro Golgi reassembly assay. In addition, mutation corresponding to the p47 F253S mutation in p37 and ufd1 also abolished their binding ability to p97. Structured summary: MINT- 7987189, MINT- 7987207, MINT- 7987303: p47 (uniprotkb:. O35987) binds (MI:. 0407) to p97 (uniprotkb:. Q01853) by pull down (MI:. 0096). MINT- 7987226: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p47 (uniprotkb:. O35987) by pull down (MI:. 0096). MINT- 7987348: p97 (uniprotkb:. P46462) physically interacts (MI:. 0915) with Ufd1 (uniprotkb:. P70362) by pull down (MI:. 0096). MINT- 7987264: p97 (uniprotkb:. P46462) and p47 (uniprotkb:. O35987) bind (MI:. 0407) by competition binding (MI:. 0405). MINT- 7987326: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p37 (uniprotkb:. Q0KL01) by pull down (MI:. 0096).

Original languageEnglish
Pages (from-to)3873-3877
Number of pages5
JournalFEBS Letters
Volume584
Issue number18
DOIs
Publication statusPublished - Sep 1 2010

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Carrier Proteins
Mutation
Assays
Membrane Fusion
Fusion reactions
Binding Sites
Membranes
Point Mutation
Experiments
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Isolation of a point-mutated p47 lacking binding affinity to p97ATPase. / Kaneko, Yayoi; Tamura, Kaori; Totsukawa, Go; Kondo, Hisao.

In: FEBS Letters, Vol. 584, No. 18, 01.09.2010, p. 3873-3877.

Research output: Contribution to journalArticle

Kaneko, Yayoi ; Tamura, Kaori ; Totsukawa, Go ; Kondo, Hisao. / Isolation of a point-mutated p47 lacking binding affinity to p97ATPase. In: FEBS Letters. 2010 ; Vol. 584, No. 18. pp. 3873-3877.
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title = "Isolation of a point-mutated p47 lacking binding affinity to p97ATPase",
abstract = "p47, a p97-binding protein, functions in Golgi membrane fusion together with p97 and VCIP135, another p97-binding protein. We have succeeded in creating p47 with a point mutation, F253S, which lacks p97-binding affinity. p47 mapping experiments revealed that p47 had two p97-binding regions and the F253S mutation occurred in the first p97-binding site. p47(F253S) could not form a complex with p97 and did not caused any cisternal regrowth in an in vitro Golgi reassembly assay. In addition, mutation corresponding to the p47 F253S mutation in p37 and ufd1 also abolished their binding ability to p97. Structured summary: MINT- 7987189, MINT- 7987207, MINT- 7987303: p47 (uniprotkb:. O35987) binds (MI:. 0407) to p97 (uniprotkb:. Q01853) by pull down (MI:. 0096). MINT- 7987226: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p47 (uniprotkb:. O35987) by pull down (MI:. 0096). MINT- 7987348: p97 (uniprotkb:. P46462) physically interacts (MI:. 0915) with Ufd1 (uniprotkb:. P70362) by pull down (MI:. 0096). MINT- 7987264: p97 (uniprotkb:. P46462) and p47 (uniprotkb:. O35987) bind (MI:. 0407) by competition binding (MI:. 0405). MINT- 7987326: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p37 (uniprotkb:. Q0KL01) by pull down (MI:. 0096).",
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N2 - p47, a p97-binding protein, functions in Golgi membrane fusion together with p97 and VCIP135, another p97-binding protein. We have succeeded in creating p47 with a point mutation, F253S, which lacks p97-binding affinity. p47 mapping experiments revealed that p47 had two p97-binding regions and the F253S mutation occurred in the first p97-binding site. p47(F253S) could not form a complex with p97 and did not caused any cisternal regrowth in an in vitro Golgi reassembly assay. In addition, mutation corresponding to the p47 F253S mutation in p37 and ufd1 also abolished their binding ability to p97. Structured summary: MINT- 7987189, MINT- 7987207, MINT- 7987303: p47 (uniprotkb:. O35987) binds (MI:. 0407) to p97 (uniprotkb:. Q01853) by pull down (MI:. 0096). MINT- 7987226: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p47 (uniprotkb:. O35987) by pull down (MI:. 0096). MINT- 7987348: p97 (uniprotkb:. P46462) physically interacts (MI:. 0915) with Ufd1 (uniprotkb:. P70362) by pull down (MI:. 0096). MINT- 7987264: p97 (uniprotkb:. P46462) and p47 (uniprotkb:. O35987) bind (MI:. 0407) by competition binding (MI:. 0405). MINT- 7987326: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p37 (uniprotkb:. Q0KL01) by pull down (MI:. 0096).

AB - p47, a p97-binding protein, functions in Golgi membrane fusion together with p97 and VCIP135, another p97-binding protein. We have succeeded in creating p47 with a point mutation, F253S, which lacks p97-binding affinity. p47 mapping experiments revealed that p47 had two p97-binding regions and the F253S mutation occurred in the first p97-binding site. p47(F253S) could not form a complex with p97 and did not caused any cisternal regrowth in an in vitro Golgi reassembly assay. In addition, mutation corresponding to the p47 F253S mutation in p37 and ufd1 also abolished their binding ability to p97. Structured summary: MINT- 7987189, MINT- 7987207, MINT- 7987303: p47 (uniprotkb:. O35987) binds (MI:. 0407) to p97 (uniprotkb:. Q01853) by pull down (MI:. 0096). MINT- 7987226: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p47 (uniprotkb:. O35987) by pull down (MI:. 0096). MINT- 7987348: p97 (uniprotkb:. P46462) physically interacts (MI:. 0915) with Ufd1 (uniprotkb:. P70362) by pull down (MI:. 0096). MINT- 7987264: p97 (uniprotkb:. P46462) and p47 (uniprotkb:. O35987) bind (MI:. 0407) by competition binding (MI:. 0405). MINT- 7987326: p97 (uniprotkb:. P46462) binds (MI:. 0407) to p37 (uniprotkb:. Q0KL01) by pull down (MI:. 0096).

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