Inactivation of annexin II tetramer by S-nitrosoglutathione.
We investigated the effect of nitric oxide (NO) donors on the activities of annexin II tetramer (AIIt), a member of the Ca2+- dependent phospholipid-binding protein family. Incubation of purified AIIt with S-nitrosoglutathione (GSNO) led to the inhibition of AIIt-mediated liposome aggregation. This effect was dose-dependent with an IC50 of approximately 100 micro m. Sodium nitroprusside, another NO donor also inhibited AIIt-mediated liposome aggregation, whereas reduced glutathione, nitrate, or nitrite had no effects. GSNO also inhibited AIIt-mediated membrane fusion, but not the binding of AIIt to the membrane. GSNO only has a modest effect on liposome aggregation mediated by annexins I, III or IV. The binding of AIIt to the membrane protected the reactive sites of GSNO on AIIt. GSNO did not inhibit AIIt-mediated liposome aggregation in the presence of dithiothreitol. Taken together, our results suggest that GSNO inactivates AIIt possibly via S-nitrosylation and/or the formation of disulfide bonds.
Department of Physiological Sciences, Oklahoma State University, Stillwater 74078, USA. liulin@okstate.edu
Lin
L
Liu
Edward
E
Enright
Peng
P
Sun
Shwu Yar
SY
Tsai
Pragna
P
Mehta
David L
DL
Beckman
David M
DM
Terrian
D000818:Animals; D017306:Annexin A2 / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002837:Chromaffin Granules / Q000502:physiology; D003545:Cysteine / Q000737:chemistry; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005033:Ethylmaleimide / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D066298:In Vitro Techniques; D008081:Liposomes; D008168:Lung / Q000737:chemistry; D008561:Membrane Fusion / Q000187:drug effects; D008567:Membranes, Artificial; D009566:Nitrates / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009573:Nitrites / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D026422:S-Nitrosoglutathione / Q000494:pharmacology*
D017306:Annexin A2; D008081:Liposomes; D008567:Membranes, Artificial; D009566:Nitrates; D020030:Nitric Oxide Donors; D009573:Nitrites; D009599:Nitroprusside; D026422:S-Nitrosoglutathione; D005978:Glutathione; D003545:Cysteine; D005033:Ethylmaleimide; D002118:Calcium; D004229:Dithiothreitol