150-kDa oxygen-regulated protein (ORP150) functions as a novel molecular chaperone in MDCK cells.
To assess the participation of the 150-kDa oxygen-regulated protein (ORP150) in protein transport, its function in Madin-Darby canine kidney (MDCK) cells was studied. Exposure of MDCK cells to hypoxia resulted in an increase of ORP150 antigen and increased binding of ORP150 to GP80/clusterin (80-kDa glycoprotein), a natural secretory protein in this cell line. In ORP150 antisense transformant MDCK cells, GP80 was retained within the endoplasmic reticulum after exposure to hypoxia. Metabolic labeling showed the delay of GP80 maturation in antisense transformants in hypoxia, whereas its matured form was detected in wild-type cells, indicating a role of ORP150 in protein transport, especially in hypoxia. The affinity chromatographic analysis of ORP150 suggested its ability to bind to ATP-agarose. Furthermore, the ATP hydrolysis analysis showed that ORP150 can release GP80 at a lower ATP concentration. These data indicate that ORP150 may function as a unique molecular chaperone in renal epithelial cells by facilitating protein transport/maturation in an environment where less ATP is accessible.
Bando
Y
Y
Department of Anatomy and Neuroscience, Osaka University Graduate School of Medicine, Suita City, Japan. ybando@anat2.med.osaka-u.ac.jp
Ogawa
S
S
Yamauchi
A
A
Kuwabara
K
K
Ozawa
K
K
Hori
O
O
Yanagi
H
H
Tamatani
M
M
Tohyama
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival; D051152:Clusterin; D004285:Dogs; D004734:Energy Metabolism; D006023:Glycoproteins / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins; D006801:Humans; D007668:Kidney; D008562:Membrane Glycoproteins / Q000378:metabolism; D018832:Molecular Chaperones / Q000502:physiology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
C495242:CLU protein, human; D051152:Clusterin; D006023:Glycoproteins; D018840:HSP70 Heat-Shock Proteins; C508603:IL6R protein, human; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D020319:Oligodeoxyribonucleotides, Antisense; D011506:Proteins; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins; C050087:oxygen-regulated proteins; D000255:Adenosine Triphosphate