title: Disintegration of lysosomes mediated by GTPgammaS-treated cytosol: possible involvement of phospholipases.
abstract: We showed previously that cytosol treated with guanosine 5'-O-(3-thiotriphosphate) (GTP-gammaS) disintegrated lysosomes in vitro [Sai, Y. et al. (1994) Biochem. Biophys. Res. Commun. 198, 869-877] in time-, temperature-, and dose-dependent manners. This also requires ATP, however, the latter can be substituted with deoxy-ATP, ADP, or ATPgammaS, suggesting no requirement of ATP hydrolysis. The lysis was inhibited by several chemical modifiers, including N-ethylmaleimide, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, and by various phospholipase inhibitors (trifluoperazine, p-bromophenacyl bromide, nordihydroguaiaretic acid, W-7, primaquine, compound 48/80, neomycin, and gentamicin), but not by ONO-RS-082, an inhibitor of phospholipase A2. The reaction was also inhibited by phospholipids (phosphatidylinositol, phosphatidylserine, phosphatidic acid, and phosphatidylcholine) and diacylglycerol. Among the phospholipase A2 hydrolysis products of phospholipids, unsaturated fatty acids (oleate, linoleate, and arachidonate) and lysophospholipid (lysophosphatidylcholine) by themselves broke lysosomes down directly, whereas saturated fatty acids (palmitate and stearate) had little effect. We found that GTPgammaS-stimulated cytosolic phospholipase A2 activity was highly sensitive to ONO-RS-082. These results suggest the participation of phospholipase(s), though not cytosolic phospholipase A2, in the GTPgammaS-dependent lysis of lysosomes.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kanazawa University, Kanazawa 920-0934.
authors.forename: Y
authors.initials: Y
authors.lastname: Sai
authors.forename: T
authors.initials: T
authors.lastname: Matsuda
authors.forename: K
authors.initials: K
authors.lastname: Arai
authors.forename: S
authors.initials: S
authors.lastname: Ohkuma
mesh_terms: D000818:Animals; D003600:Cytosol / Q000187:drug effects* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005227:Fatty Acids / Q000494:pharmacology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology*; D008099:Liver / Q000648:ultrastructure; D008247:Lysosomes / Q000187:drug effects* / Q000378:metabolism; D008561:Membrane Fusion; D010740:Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010743:Phospholipids / Q000494:pharmacology; D051381:Rats
chemical_list: D004791:Enzyme Inhibitors; D005227:Fatty Acids; D010743:Phospholipids; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D010740:Phospholipases