Identification of ER membrane targeting signal of kinectin.
Kinectin has been identified as a kinesin receptor on endoplasmic reticulum (ER). The ER membrane binding domain of kinectin is still obscure and is thought to require a half of the molecule. To determine the ER insertion site, we produced several constructs around N-terminus of kinectin connected with green fluorescent protein (GFP) and visualized the distribution in Cos-7 cells. The fragment of residues 7-29 appeared in the reticular pattern exactly colocalized with the ER marker but did not remain for a long time. On the other hand, residues 1-106 maintained a reticular pattern for more than seven days. These results indicate that residues 7-29 of kinectin are sufficient for targeting to the ER membrane but insufficient for remaining on the ER.
Abe
Erika
E
Department of Neurology, Akita University School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan. erikaa@doc.med.akita-u.ac.jp
Okawa
Satoshi
S
Sugawara
Masashiro
M
Watanabe
Sumio
S
Toyoshima
Itaru
I
D000818:Animals; D001692:Biological Transport; D019556:COS Cells / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014162:Transfection / Q000379:methods
C088547:KTN1 protein, human; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins