Dataset Information

Autologous extracellular cytochrome c is an endogenous ligand for leucine-rich alpha2-glycoprotein and beta-type phospholipase A2 inhibitor.

ABSTRACT: Beta-type phospholipase A(2) inhibitory protein (PLIbeta) from the serum of the venomous snake Gloydius brevicaudus neutralizes basic phospholipase A(2) (PLA(2)) from its own venom, and it has 33% sequence homology with human leucine-rich alpha(2)-glycoprotein (LRG), which has been recently reported to bind cytochrome c (Cyt c) (Cummings, C., Walder, J., Treeful, A., and Jemmerson, R. (2006) Apoptosis 11, 1121-1129). In the present study, PLIbeta was found to bind Cyt c. The interactions of LRG and PLIbeta with Cyt c were compared by surface plasmon resonance analysis. Human LRG bound horse and snake Cyt c with dissociation constants of 1.58 x 10(-13) M and 1.65 x 10(-10) M, respectively, but did not bind yeast Cyt c, while G. brevicaudus PLIbeta bound horse, snake, and yeast Cyt c with dissociation constants of 1.05 x 10(-10) M, 2.37 x 10(-12) M, and 1.67 x 10(-6) M, respectively. On the other hand, LRG did not show any PLA(2) inhibitory activity and did not bind G. brevicaudus basic PLA(2), whereas PLIbeta bound the basic PLA(2) with a dissociation constant of 1.21 x 10(-9) M, which is smaller than those with the Cyt c described above. The PLA(2) inhibitory activity of PLIbeta was also found to be suppressed by the binding of Cyt c to PLIbeta. These results suggest that autologous Cyt c is an endogeneous ligand for LRG and PLIbeta and that these serum proteins neutralize the autologous Cyt c released from the dead cells.

SUBMITTER: Shirai R

PROVIDER: S-EPMC2898406 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Autologous extracellular cytochrome c is an endogenous ligand for leucine-rich alpha2-glycoprotein and beta-type phospholipase A2 inhibitor.

Shirai Ryoichi R Gotou Rikiya R Hirano Fumiyasu F Ikeda Kiyoshi K Inoue Seiji S

The Journal of biological chemistry 20100504 28

Beta-type phospholipase A(2) inhibitory protein (PLIbeta) from the serum of the venomous snake Gloydius brevicaudus neutralizes basic phospholipase A(2) (PLA(2)) from its own venom, and it has 33% sequence homology with human leucine-rich alpha(2)-glycoprotein (LRG), which has been recently reported to bind cytochrome c (Cyt c) (Cummings, C., Walder, J., Treeful, A., and Jemmerson, R. (2006) Apoptosis 11, 1121-1129). In the present study, PLIbeta was found to bind Cyt c. The interactions of LRG ...[more]

PMID: 20442399

Similar Datasets

Project description:BackgroundPhospholipase A2s mediate the rate-limiting step in the formation of eicosanoids such as cysteinyl leukotrienes (CysLTs). Group IVA cytosolic PLA2α (cPLA2α) is thought to be the dominant PLA2 in eosinophils; however, eosinophils also have secreted PLA2 (sPLA2) activity that has not been fully defined.ObjectivesTo examine the expression of sPLA2 group X (sPLA2-X) in eosinophils, the participation of sPLA2-X in the formation of CysLTs, and the mechanism by which sPLA2-X initiates the synthesis of CysLTs in eosinophils.MethodsPeripheral blood eosinophils were obtained from volunteers with asthma and/or allergy. A rabbit polyclonal anti-sPLA2-X antibody identified sPLA2-X by Western blot. We used confocal microscopy to colocalize the sPLA2-X to intracellular structures. An inhibitor of sPLA2-X (ROC-0929) that does not inhibit other mammalian sPLA2s, as well as inhibitors of the mitogen-activated kinase cascade (MAPK) and cPLA2α, was used to examine the mechanism of N-formyl-methionyl-leucyl-phenylalanine (fMLP)-mediated formation of CysLT.ResultsEosinophils express the mammalian sPLA2-X gene (PLA2G10). The sPLA2-X protein is located in the endoplasmic reticulum, golgi, and granules of eosinophils and moves to the granules and lipid bodies during fMLP-mediated activation. Selective sPLA2-X inhibition attenuated the fMLP-mediated release of arachidonic acid and CysLT formation by eosinophils. Inhibitors of p38, extracellular-signal-regulated kinases 1/2 (p44/42 MAPK), c-Jun N-terminal kinase, and cPLA2α also attenuated the fMLP-mediated formation of CysLT. The sPLA2-X inhibitor reduced the phosphorylation of p38 and extracellular-signal-regulated kinases 1/2 (p44/42 MAPK) as well as cPLA2α during cellular activation, indicating that sPLA2-X is involved in activating the MAPK cascade leading to the formation of CysLT via cPLA2α. We further demonstrate that sPLA2-X is activated before secretion from the cell during activation. Short-term priming with IL-13 and TNF/IL-1β increased the expression of PLA2G10 by eosinophils.ConclusionsThese results demonstrate that sPLA2-X plays a significant role in the formation of CysLTs by human eosinophils. The predominant role of the enzyme is the regulation of MAPK activation that leads to the phosphorylation of cPLA2α. The sPLA2-X protein is regulated by proteolytic cleavage, suggesting that an inflammatory environment may promote the formation of CysLTs through this mechanism. These results have important implications for the treatment of eosinophilic disorders such as asthma.

Dataset Information

Autologous extracellular cytochrome c is an endogenous ligand for leucine-rich alpha2-glycoprotein and beta-type phospholipase A2 inhibitor.

Publications

Autologous extracellular cytochrome c is an endogenous ligand for leucine-rich alpha2-glycoprotein and beta-type phospholipase A2 inhibitor.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets