Isolation of a point-mutated p47 lacking binding affinity to p97ATPase.
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.
Kaneko
Yayoi
Y
Department of Molecular Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Tamura
Kaori
K
Totsukawa
Go
G
Kondo
Hisao
H
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D018797:Cell Cycle Proteins; D010450:Endopeptidases / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D017354:Point Mutation; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism; D051381:Rats; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000235:genetics / Q000378:metabolism*
D018797:Cell Cycle Proteins; C498751:Nsfl1c protein, rat; D009687:Nuclear Proteins; D011506:Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; C491496:Ufd1 protein, rat; D010450:Endopeptidases; C470473:Vcpip1 protein, rat; D000251:Adenosine Triphosphatases; C404942:p97 ATPase