Identification of a novel, Ca2+‐dependent phospholipase D with preference for phosphatidylserine and phosphatidylethanolamine in Saccharomyces cerevisiae

FEBS Letters

Paltauf, Fritz¹  Mayr, Johannes A.¹  Kohlwein, Sepp D.¹ 

[1] Institut für Biochemie und Lebensmittelchemie, Technische Universität Graz, Petersgasse 12/II, A-8010 Graz, Austria

关键词: Phospholipase D;    Saccharomyces cerevisiae;    Yeast;    Phospholipid;    Transphosphatidylation;    PLD1;    Calcium;    Signalling;    PLD;    phospholipase D (EC 3.1.4.4);    Ca-PLD;    Ca2+-dependent phospholipase D;    Pld1p;    product of the PLD1 gene;    PKC;    protein kinase C;    PtdCho;    phosphatidylcholine;    PtdEtn;    phosphatidylethanolamine;    PtdSer;    phosphatidylserine;    PtdIns;    phosphatidylinositol;    PtdOH;    phosphatidic acid;    PtdEt;    phosphatidylethanol;    NBD-PtdCho;    1-palmitoyl-2-[6-N-(7-nitro-2-oxo-1;    3-benzodiazol-4-yl)-amino]caproyl-sn-glycero-3-phosphocholine;    PIP2;    phosphatidylinositol-4;    5-bisphosphate;    PMSF;    phenylmethanesulfonylfluoride;    EDTA;    ethylenediaminetetraacetic acid;    PM;    plasma membrane;   

DOI  :  10.1016/0014-5793(96)00893-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A membrane-bound phospholipase D (PLD) from Saccharomyces cerevisiae was solubilized from mitochondrial and plasma membranes and partially purified. The enzyme has an apparent molecular weight of approximately 60 kDa, is strictly Ca²⁺-dependent and preferentially hydrolyses phosphatidylserine and phosphatidylethanolamine. Enzyme activity is significantly increased in membranes from cells grown on a non-fermentable carbon source. The Ca²⁺-dependent PLD is distinct from PLD encoded by the SPO14/PLD1 gene. The 195 kDa SPO14/PLD1 gene product is specific for PtdCho, Ca²⁺-independent and is activated by PIP₂. Furthermore, Pld1p has transphosphatidylation activity in the presence of ethanol and thus resembles the prototypic PLD activity found in mammalian cells and plants. In contrast, the Ca²⁺-dependent PLD described here is not affected by PIP₂ and does not catalyze transphosphatidylation. Thus, the Ca²⁺-dependent PLD characterized in this study appears to be a member of a novel family of phospholipases D.

【 授权许可】

Unknown