title: Phospholipase D2 localizes to the plasma membrane and regulates angiotensin II receptor endocytosis.
abstract: Phospholipase D (PLD) is a key facilitator of multiple types of membrane vesicle trafficking events. Two PLD isoforms, PLD1 and PLD2, exist in mammals. Initial studies based on overexpression studies suggested that in resting cells, human PLD1 localized primarily to the Golgi and perinuclear vesicles in multiple cell types. In contrast, overexpressed mouse PLD2 was observed to localize primarily to the plasma membrane, although internalization on membrane vesicles was observed subsequent to serum stimulation. A recent report has suggested that the assignment of PLD2 to the plasma membrane is in error, because the endogenous isoform in rat secretory cells was imaged and found to be present primarily in the Golgi apparatus. We have reexamined this issue by using a monoclonal antibody specific for mouse PLD2, and find, as reported initially using overexpression studies, that endogenous mouse PLD2 is detected most readily at the plasma membrane in multiple cell types. In addition, we report that mouse, rat, and human PLD2 when overexpressed all similarly localize to the plasma membrane in cell lines from all three species. Finally, studies conducted using overexpression of wild-type active or dominant-negative isoforms of PLD2 and RNA interference-mediated targeting of PLD2 suggest that PLD2 functions at the plasma membrane to facilitate endocytosis of the angiotensin II type 1 receptor.
authors.affiliation: Department of Pharmacology and the Center for Developmental Genetics, University Medical Center at Stony Brook, Stony Brook, New York 11794-5140, USA.
authors.forename: Guangwei
authors.initials: G
authors.lastname: Du
authors.forename: Ping
authors.initials: P
authors.lastname: Huang
authors.forename: Bruce T
authors.initials: BT
authors.lastname: Liang
authors.forename: Michael A
authors.initials: MA
authors.lastname: Frohman
mesh_terms: D041721:3T3-L1 Cells; D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002462:Cell Membrane / Q000201:enzymology*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis / Q000502:physiology*; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D051379:Mice; D032342:Muscle Cells / Q000201:enzymology; D009154:Mutation / Q000235:genetics; D009206:Myocardium / Q000201:enzymology; D010739:Phospholipase D / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D011945:Receptors, Angiotensin / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D034741:RNA, Small Interfering; D011945:Receptors, Angiotensin; C105606:phospholipase D2; D010739:Phospholipase D