TY - JOUR
T1 - Isolation of a point-mutated p47 lacking binding affinity to p97ATPase
AU - Kaneko, Yayoi
AU - Tamura, Kaori
AU - Totsukawa, Go
AU - Kondo, Hisao
N1 - Funding Information:
We would like to thank H.A. Popiel for her kind assistance in preparation of the manuscript. This work is supported by a grant to H.K. from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
PY - 2010/9
Y1 - 2010/9
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).
UR - http://www.scopus.com/inward/record.url?scp=77956943702&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77956943702&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2010.07.061
DO - 10.1016/j.febslet.2010.07.061
M3 - Article
C2 - 20691684
AN - SCOPUS:77956943702
SN - 0014-5793
VL - 584
SP - 3873
EP - 3877
JO - FEBS Letters
JF - FEBS Letters
IS - 18
ER -