title: Acetylcholinesterase conformational states influence nitric oxide mobilization in the erythrocyte.
abstract: In the human erythrocyte, band 3 protein mediates nitric oxide (NO) translocation and its effects are strongly related to phosphorylated/dephosphorylated intracellular states. The metabolism of NO could change in the presence of acetylcholinesterase (AChE). Therefore, the present study was designed to assess the effect of conformational changes in AChE (via N-19 and C-16 antibodies) and enzymatic inhibition/activation of protein kinase C (PKC) in erythrocyte NO mobilization in vitro. Our results show that by inhibiting PKC with cheletrine, impaired erythrocyte NO efflux and s-nitrosoglutathione (GSNO) levels were verified, while PKC's activation by Phorbol 12-myristate 13-acetate had the opposite effect. Those results demonstrate the influence of 4.1R complex and band 3 protein level of phosphorylation on NO efflux and GSNO concentration mediated by PKC inhibition/activation. In addition, the present study shows evidence that conformational changes in AChE promoted by incubation with N-19 and C-16 antibodies alter the enzyme's functional connection to acetylcholine (ACh) (AChE-ACh complex) in an irreversible manner, resulting in impaired GSNO concentration and NO efflux from the erythrocyte. Novel insight into NO metabolism in the erythrocyte is brought with the presented findings allowing new possibilities of modulating NO delivery, possibly involving PKC and AChE conformational alterations in combination.
authors: Carlota Saldanha Lab, Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Edifício Egas Moniz, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
authors: Pedro
authors: P
authors: Teixeira
authors: Nuno
authors: N
authors: Duro
authors: Patrícia
authors: P
authors: Napoleão
authors: Carlota
authors: C
authors: Saldanha
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000110:Acetylcholinesterase / Q000737:chemistry* / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000737:chemistry / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism*; D011487:Protein Conformation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D026422:S-Nitrosoglutathione / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D011493:Protein Kinase C; D000110:Acetylcholinesterase; D000109:Acetylcholine
references: Nature. 1981 Sep 10-16;293(5828):144-6
references: 7266668
references: Clin Hemorheol Microcirc. 2008;40(3):207-27
references: 19029645
references: Biochem Biophys Res Commun. 1995 Aug 4;213(1):249-57
references: 7543753
references: Biosens Bioelectron. 2004 Oct 15;20(3):505-8
references: 15494232
references: J Cereb Blood Flow Metab. 2014 Apr;34(4):613-20
references: 24447953
references: Eur J Biochem. 1999 Sep;264(3):672-86
references: 10491113
references: Cell Mol Neurobiol. 1991 Feb;11(1):157-72
references: 1849452
references: J Membr Biol. 2009 Sep;231(1):47-53
references: 19809847
references: Biochem J. 1993 Dec 15;296 ( Pt 3):675-83
references: 8280066
references: Anal Biochem. 1996 Jul 1;238(2):150-8
references: 8660604
references: Biochem Biophys Res Commun. 1990 Nov 15;172(3):993-9
references: 2244923
references: Biochem J. 2000 Oct 1;351(Pt 1):95-105
references: 10998351
references: J Biol Chem. 1986 Feb 25;261(6):2804-9
references: 3949748
references: Clin Hemorheol Microcirc. 2013;54(4):357-69
references: 23076000
references: Nature. 1980 Nov 27;288(5789):373-6
references: 6253831
references: J Biol Chem. 1998 Jul 31;273(31):19829-33
references: 9677417
references: Int J Cardiol. 2014 Jun 15;174(2):230-42
references: 24794552
references: Biochem J. 1973 Jul;133(3):521-7
references: 4733238
references: Clin Hemorheol Microcirc. 2012;51(1):1-20
references: 22240380
references: Clin Hemorheol Microcirc. 2008;39(1-4):363-73
references: 18503146
references: Biochim Biophys Acta. 1991 Jan 30;1061(2):253-66
references: 1998697
references: Nat Rev Mol Cell Biol. 2010 Feb;11(2):103-12
references: 20094051
references: Nat Biotechnol. 2011 Oct 30;29(11):1039-45
references: 22037377
references: J Membr Biol. 2009 Mar;228(2):89-97
references: 19294450
references: Clin Exp Pharmacol Physiol. 1999 Mar;26(3):198-205
references: 10081614
references: J Appl Toxicol. 2004 Nov-Dec;24(6):419-27
references: 15551380
references: J Biol Chem. 2000 Mar 3;275(9):6360-7
references: 10692436
references: J Biol Chem. 2011 Aug 19;286(33):28922-30
references: 21715334
references: Biochim Biophys Acta. 1980 Mar 14;612(1):67-72
references: 7362833
references: Int J Mol Sci. 2012;13(9):10697-721
references: 23109817
references: J Mol Neurosci. 2006;30(1-2):233-6
references: 17192683
references: Pharmaceuticals (Basel). 2013;6(3):407-39
references: 23580870
references: Front Biosci. 2004 Sep 01;9:2063-85
references: 15353271