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Activation of Phospholipase D2 through Phosphorylation of Tyrosine-470 in Antigen-stimulated Mast Cells

  • Published : 2005.06.01

Abstract

The mechanism of activation of phospholipase D2 (PLD2) remains undefined although mechanisms have been described for the activation of PLDI. By expression of mutated forms of haemaglutinnin-tagged PLD2 in a mast cell (RBL-2H3) line, we show that PLD2 is phosphorylated at tyrosines -11, -14, and -470 and that tyrosine-470 is critical for activation of PLD2 by antigen. Studies were performed with mutated-DNA constructs for haemaglutinnin-tagged PLD2 in which codons for tyrosine -11, -14, -165, and -470 were mutated to phenylalanine either individually or collectively. Transient expression of these constructs showed that mutation of tyrosine -11, -14, -470, or all tyrosines (all-mutated PLD2) suppressed antigen-induced tyrosine phosphorylation of PLD2 but only the tyrosine-470 mutant failed to be activated by antigen as assessed by in vitro assay of immunoprepitated PLD2 or by assay of PLD in intact cells. The critical role of tyrosine-470 was confirmed in studies with add-back mutants (phenylalanine back to tyrosine) of the all-mutated PLD. The findings provide the first description of a mechanism of activation of PLD2 in a physiological setting.

PLDI 활성화 기전은 여러 보고가 있으나 PLD2 활성화에 대한 기전은 아직 연구의 대상이다. RBL-2H3 비만세포에서 HA-PLD2의 인산화 가능한 타이로신 잔기를 점돌연변이 시킨 DNA플라즈미드를 이용하여 11번, 14번, 470번의 타이로신이 항원자극에 의해 인산화 됨을 알아냈고 특히 470번 타이로신의 인산화가 PLD2 활성화에 중요하다는 결과를 얻었다.

Keywords

References

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