Sung Ho Ryu

Two forms of phosphatidylinositol-specific phospholipase C from bovine brain.

Two immunologically distinct forms of phosphoinositide-specific phospholipase C were purified to near homogeneity from bovine brain. Their molecular weights determined by SDS-PAGE are 150,000 (enzyme I) and 145,000 (enzyme II), respectively. Under a nondenaturing condition, purified enzyme I exists mainly in dimeric form and as tetramer to a small extent, while enzyme II is predominantly in monomer, to a small extent as dimer and to a very small extent as trimer. Multiple forms of phosphoinositide-specific phospholipase C in brain tissue described in the literature might be, therefore attributed to the oligomerization of the two independent forms.

Cyclic and noncyclic inositol phosphates are formed at different ratios by phospholipase C isozymes

The cyclic inositol phosphate content in the product of PLC-beta, gamma, and delta mediated cleavage of three phosphoinositides, PtdIns, PtdIns(4)p, and PtdIns(4,5)P2, was measured under several different experimental conditions. The ratio of cyclic to noncyclic product generally decreased in the order PLC-beta greater than PLC-delta greater than PLC-gamma. For all three enzymes the ratio decreased in the order PtdIns greater than PtdIns(4)P greater than PtdIns(4,5)P2. For all combinations of the three enzymes and three substrates cyclic product content was always higher at pH 5.5 than at pH 7.0. The effect of Ca2+ on the ratio of cyclic to noncyclic was also measured. The ratio remained constant between 0.5 microM and 2 mM for PtdIns. For PtdIns(4)P and PtdIns(4,5)P2, the ratios were unchanged between 0.5 and 500 microM, but increased abruptly at millimolar Ca2+ concentrations.

OGA heterozygosity suppresses intestinal tumorigenesis in Apc min/+ mice

Emerging evidence suggests that aberrant O-GlcNAcylation is associated with tumorigenesis. Many oncogenic factors are O-GlcNAcylated, which modulates their functions. However, it remains unclear how O-GlcNAcylation and O-GlcNAc cycling enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), affect the development of cancer in animal models. In this study, we show that reduced level of OGA attenuates colorectal tumorigenesis induced by Adenomatous polyposis coli (Apc) mutation. The levels of O-GlcNAcylation and O-GlcNAc cycling enzymes were simultaneously upregulated in intestinal adenomas from mice, and in human patients. In two independent microarray data sets, the expression of OGA and OGT was significantly associated with poor cancer-specific survival of colorectal cancer (CRC) patients. In addition, OGA heterozygosity, which results in increased levels of O-GlcNAcylation, attenuated intestinal tumor formation in the Apcmin/+ background. Apcmin/+ OGA+/− mice exhibited a significantly increased survival rate compared with Apcmin/+ mice. Consistent with this, Apcmin/+ OGA+/− mice expressed lower levels of Wnt target genes than Apcmin/+. However, the knockout of OGA did not affect Wnt/β-catenin signaling. Overall, these findings suggest that OGA is crucial for tumor growth in CRC independently of Wnt/β-catenin signaling.