tailieunhanh - Báo cáo khoa học: Effect of priming on activation and localization of phospholipase D-1 in human neutrophils

Tham khảo luận văn - đề án 'báo cáo khoa học: effect of priming on activation and localization of phospholipase d-1 in human neutrophils', luận văn - báo cáo, báo cáo khoa học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Eur. J. Biochem. 271 2755-2764 2004 FEBS 2004 doi Effect of priming on activation and localization of phospholipase D-1 in human neutrophils Karen A. Cadwallader1 Mohib Uddin1 Alison M. Condliffe1 Andrew S. Cowburn1 Jessica F. White1 Jeremy N. Skepper2 Nicholas T. Ktistakis3 and Edwin R. Chilvers1 1 Respiratory Medicine Division Department of Medicine University of Cambridge School of Clinical Medicine Addenbrooke s and Papworth Hospitals Cambridge UK department of Anatomy University of Cambridge UK 3 Department of Signalling Babraham Institute Cambridge UK Phospholipase D PLD plays a major role in the activation of the neutrophil respiratory burst. However the repertoire of PLD isoforms present in these primary cells the precise mechanism of activation and the mi pact of Olli priming on PLD activity and localization remain poorly defined. RT-PCR analysis showed that both PL DI and LLD2 isoforms are expressed m Umnm neutoophils whh LDD1 expressed at a fl igher level. Endoeenuus LLD1 was deteeted by immunoprecipitation and Western l loiéog. ndd was predominantly membrane-associated under control and primed stimulated conditions. Immunofluorescence showed that PLD hada punctate distribution throuihout the cell which was not alteredafter stimulation by soluble aionists. In contrast PLD localizedto the phaiolysosome membrane after ingestion of nonopsonized bymoren anilides. We also demonstrate that tumour necrosis factor a greatly potentiates aionist-stimulatedPLD activation myelo peroxidase release and superoxide anion generation and that PLD activation occurs via a phosphatidylinositol 3-kinase-sensitive and bfeftldin-sensilive ADP-siboryPltion factor GTPase-regulated 11 011111 111. MolPOher. propranolol which causes an increase in PLD-derived phosphatidic acidaccumulation causeda selective increase in agonist-stimulated myeloperoxidase release. Our results indicate that priming is a critical regulator of PLD activation that the .