title: Inhalational anesthetics disrupt postsynaptic density protein-95, Drosophila disc large tumor suppressor, and zonula occludens-1 domain protein interactions critical to action of several excitatory receptor channels related to anesthesia.
abstract: BACKGROUND
  The authors have shown previously that inhaled anesthetics disrupt the interaction between the second postsynaptic density protein-95, Drosophila disc large tumor suppressor, and zonula occludens-1 (PDZ) domain of postsynaptic density protein-95 (PSD-95) and the C-terminus of N-methyl-D-aspartate receptor subunits NR2A and NR2B. The study data indicate that PDZ domains may serve as a molecular target for inhaled anesthetics. However, the underlying molecular mechanisms remain to be illustrated.
  
  
  METHODS
  Glutathione S-transferase pull-down assay, coimmunoprecipitation, and yeast two-hybrid analysis were used to assess PDZ domain-mediated protein-protein interactions in different conditions. Nuclear magnetic resonance spectroscopy was used to investigate isoflurane-induced chemical shift changes in the PDZ1-3 domains of PSD-95. A surface plasmon resonance-based BIAcore (Sweden) assay was used to examine the ability of isoflurane to inhibit the PDZ domain-mediated protein-protein interactions in real time.
  
  
  RESULTS
  Halothane and isoflurane dose-dependently inhibited PDZ domain-mediated interactions between PSD-95 and Shaker-type potassium channel Kv1.4 and between α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunit GluA2 and its interacting proteins-glutamate receptor-interacting protein or protein interacting with c kinase 1. However, halothane and isoflurane had no effect on PDZ domain-mediated interactions between γ-aminobutyric acid type B receptor and its interacting proteins. The inhaled anesthetic isoflurane mostly affected the residues close to or in the peptide-binding groove of PSD-95 PDZ1 and PDZ2 (especially PDZ2), while barely affecting the peptide-binding groove of PSD-95 PDZ3.
  
  
  CONCLUSIONS
  These results suggest that inhaled anesthetics interfere with PDZ domain-mediated protein-protein interactions at several receptors important to neuronal excitation, anesthesia, and pain processing.
authors: From the Department of Anesthesiology and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland (F.T., Y.S., J.S., R.A.J.); Department of Biomedical Sciences, Texas A&M University Baylor College of Dentistry, Dallas, Texas (F.T.); and Departments of Anesthesiology (Q.C., P.T.), Pharmacology and Chemical Biology (P.T.), and Computational Biology (P.T.), University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
authors: Feng
authors: F
authors: Tao
authors: Qiang
authors: Q
authors: Chen
authors: Yuko
authors: Y
authors: Sato
authors: John
authors: J
authors: Skinner
authors: Pei
authors: P
authors: Tang
authors: Roger A
authors: RA
authors: Johns
mesh_terms: D000595:Amino Acid Sequence; D018685:Anesthetics, Inhalation / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D000076122:Disks Large Homolog 4 Protein; D004305:Dose-Response Relationship, Drug; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017470:Receptors, Glutamate / Q000378:metabolism*; D000074407:SAP90-PSD95 Associated Proteins; D062826:Zonula Occludens-1 Protein / Q000378:metabolism*
chemical_list: D018685:Anesthetics, Inhalation; D000076122:Disks Large Homolog 4 Protein; C501449:Dlg4 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D017470:Receptors, Glutamate; D000074407:SAP90-PSD95 Associated Proteins; D062826:Zonula Occludens-1 Protein
references: Nature. 1994 Feb 17;367(6464):607-14
references: 7509043
references: J Biol Chem. 1993 Mar 5;268(7):4580-3
references: 7680343
references: Nature. 1995 Nov 2;378(6552):85-8
references: 7477295
references: Cell. 1996 Jun 28;85(7):1067-76
references: 8674113
references: Neuron. 1996 Aug;17(2):255-65
references: 8780649
references: Neuron. 1996 Oct;17(4):759-67
references: 8893032
references: Curr Opin Neurobiol. 1997 Jun;7(3):368-73
references: 9232802
references: Pharmacol Rev. 1997 Dec;49(4):343-67
references: 9443162
references: J Biol Chem. 1998 Jul 31;273(31):19518-24
references: 9677374
references: J Neurosci. 1998 Dec 1;18(23):9716-26
references: 9822732
references: Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):435-40
references: 9892651
references: Toxicol Lett. 1998 Nov 23;100-101:1-8
references: 10049127
references: Toxicol Lett. 1998 Nov 23;100-101:185-91
references: 10049141
references: J Neurosci. 1999 May 15;19(10):3926-34
references: 10234023
references: Science. 1999 Jun 11;284(5421):1845-8
references: 10364559
references: Trends Pharmacol Sci. 2005 Oct;26(10):503-10
references: 16126282
references: Curr Opin Anaesthesiol. 2007 Aug;20(4):300-6
references: 17620835
references: Nat Rev Neurosci. 2008 May;9(5):370-86
references: 18425091
references: Anesthesiology. 2008 May;108(5):882-7
references: 18431124
references: Nature. 2008 Aug 14;454(7206):903-6
references: 18704088
references: Mol Ther. 2008 Nov;16(11):1776-82
references: 18781143
references: J Phys Chem B. 2008 Nov 13;112(45):14312-8
references: 18821786
references: Biochim Biophys Acta. 2013 Feb;1828(2):398-404
references: 23000369
references: J Biol Chem. 2013 Dec 13;288(50):35793-800
references: 24194515
references: Trends Cell Biol. 2000 Jul;10(7):274-80
references: 10856930
references: J Exp Med. 2000 Dec 18;192(12):1697-706
references: 11120767
references: Annu Rev Neurosci. 2001;24:1-29
references: 11283303
references: J Biol Chem. 2001 Apr 20;276(16):12974-82
references: 11150294
references: Anesthesiology. 2001 Jun;94(6):1010-5
references: 11465592
references: Neuroreport. 2001 Oct 29;12(15):3251-5
references: 11711866
references: J Biol Chem. 2002 Mar 1;277(9):6967-73
references: 11744724
references: Neuroscience. 2003;117(3):731-9
references: 12617977
references: Sci STKE. 2003 Apr 22;2003(179):RE7
references: 12709532
references: N Engl J Med. 2003 May 22;348(21):2110-24
references: 12761368
references: J Biol Chem. 2003 Sep 19;278(38):36669-75
references: 12853458
references: Nat Rev Neurosci. 2004 Sep;5(9):709-20
references: 15322529
references: J Biomol NMR. 2004 Sep;30(1):111-2
references: 15452444
references: Br J Anaesth. 1971 Oct;43(10):913-8
references: 5115029
references: Science. 1982 Sep 10;217(4564):1055-7
references: 7112112
references: Acta Physiol Scand. 1987 Aug;130(4):679-85
references: 3630740
references: Science. 1989 Apr 14;244(4901):221-4
references: 2539643
references: Methods Enzymol. 1989;176:279-311
references: 2811690
references: Ann N Y Acad Sci. 1991;625:532-9
references: 1905502
references: J Neurophysiol. 1991 Dec;66(6):2014-21
references: 1667416
references: Proc Natl Acad Sci U S A. 1992 May 15;89(10):4329-32
references: 1584767
references: Neuron. 1992 Nov;9(5):929-42
references: 1419001
references: Science. 1995 Sep 22;269(5231):1737-40
references: 7569905