Overexpression of plasma membrane annexin II in NO2-exposed pulmonary artery endothelial cells.
Because exposure to nitrogen dioxide (NO2) alters plasma membrane structure and function in pulmonary artery endothelial cells (PAEC), we examined whether NO2 exposure is associated with upregulation of plasma membrane-specific proteins in PAEC. Exposure to 5 ppm NO2 for 24 h had no significant effect on total protein synthesis. However, two-dimensional gel electrophoresis of isolated plasma membranes from [35S]-methionine pulse-labeled PAEC exposed to NO2 for 24 h demonstrated 3- to 9-fold increases in the synthesis of several proteins with molecular masses of 36, 39, and 40 kDa compared with controls. N-terminal amino acid sequencing and immunodetection analysis identified the 36kDa plasma membrane protein as annexin II (lipocortin II). Northern blotting analysis demonstrated that the mRNA expression for annexin II in NO2-exposed cells was also increased. These results suggest that exposure to NO2 results in induction of plasma membrane annexin II, an important multifunctional calcium- and phospholipid-binding protein in PAEC.
Li
Y D
YD
Division of Pulmonary Medicine, University of Florida College of Medicine, Gainesville, USA.
Patel
J M
JM
Zhang
J
J
Block
E R
ER
D000595:Amino Acid Sequence; D000818:Animals; D017306:Annexin A2 / Q000096:biosynthesis* / Q000235:genetics; D015152:Blotting, Northern; D015153:Blotting, Western; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism*; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009585:Nitrogen Dioxide / Q000494:pharmacology*; D011651:Pulmonary Artery; D012333:RNA, Messenger / Q000378:metabolism; D017421:Sequence Analysis; D013552:Swine; D015854:Up-Regulation / Q000502:physiology
D017306:Annexin A2; D008565:Membrane Proteins; D012333:RNA, Messenger; D009585:Nitrogen Dioxide