Novel evidence of expression and activity of ecto-phospholipase C gamma1 in human T lymphocytes.
Although much is known about the intracellular phospholipase C (PLC) specific for inositol phospholipids, few data are available about the presence of a less common PLC at the external side of the membrane bilayer of some cell types. This ectoenzyme seems to play particular roles in cellular function by hydrolyzing inositol lipids located on the outer leaflet of the plasma membrane. Here, we provide the first evidence that peripheral T lymphocytes express a discrete level of a PLCgamma1 at the outer leaflet of the plasma membrane. Flow cytometry showed that the PLCgamma1-positive (PLCgamma1(+)) cells (approximately 37%) were CD8(+) and CD45RA+. Biochemical evidence indicated that (1) this ectoenzyme displays a mass similar to the cytoplasmic form, (2) it is phosphorylated on tyrosine residues, and (3) its activity is Ca2+-dependent. In addition, this enzyme appeared to be correlated with the proliferative state of the cell, since stimulation with phytohemagglutinin (PHA) downregulated both its expression and activity, which were restored by treatment with an antiproliferative agent like natural interferon beta. Moreover, the different kinetics of formation of its hydrolytic products, inositol 1 phosphate and inositol 1:2 cyclic phosphate (Ins(1)P and Ins(1:2 cycl)P), formed upon incubation of the lymphocytes with [3H]-lyso-phosphatidylinositol (PI), allow the hypothesis of a selective involvement of the two inositol phosphates in the mechanisms regulating the metabolism of particular T-lymphocyte subsets.
Miscia
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Istituto di Morfologia Umana Normale and Cattedra di Neonatologia, Università G. D'Annunzio, Chieti, Italy.
Di Baldassarre
A
A
Cataldi
A
A
Rana
R A
RA
Di Valerio
V
V
Sabatino
G
G
D000328:Adult; D002118:Calcium / Q000502:physiology; D002455:Cell Division; D002462:Cell Membrane / Q000201:enzymology*; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000096:biosynthesis; D016899:Interferon-beta / Q000494:pharmacology; D007527:Isoenzymes / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D008213:Lymphocyte Activation / Q000187:drug effects; D008246:Lysophospholipids / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism*; D018613:Microscopy, Confocal; D010716:Phosphatidylinositols / Q000378:metabolism*; D051966:Phospholipase C gamma; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis; D010835:Phytohemagglutinins / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000201:enzymology*; D010738:Type C Phospholipases / Q000096:biosynthesis / Q000378:metabolism*
D007295:Inositol Phosphates; D007527:Isoenzymes; D008246:Lysophospholipids; D008563:Membrane Lipids; D010716:Phosphatidylinositols; D010835:Phytohemagglutinins; C025449:lysophosphatidylinositol; D019000:Phosphotyrosine; D016899:Interferon-beta; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D002118:Calcium