The N-terminal domain of Drosophila Gram-negative binding protein 3 (GNBP3) defines a novel family of fungal pattern recognition receptors.
Gram-negative binding protein 3 (GNBP3), a pattern recognition receptor that circulates in the hemolymph of Drosophila, is responsible for sensing fungal infection and triggering Toll pathway activation. Here, we report that GNBP3 N-terminal domain binds to fungi upon identifying long chains of beta-1,3-glucans in the fungal cell wall as a major ligand. Interestingly, this domain fails to interact strongly with short oligosaccharides. The crystal structure of GNBP3-Nter reveals an immunoglobulin-like fold in which the glucan binding site is masked by a loop that is highly conserved among glucan-binding proteins identified in several insect orders. Structure-based mutagenesis experiments reveal an essential role for this occluding loop in discriminating between short and long polysaccharides. The displacement of the occluding loop is necessary for binding and could explain the specificity of the interaction with long chain structured polysaccharides. This represents a novel mechanism for beta-glucan recognition.
Mishima
Yumiko
Y
Centre de Biophysique Moléculaire, UPR 4301 CNRS, 45071 Orléans Cedex 2, France.
Quintin
Jessica
J
Aimanianda
Vishukumar
V
Kellenberger
Christine
C
Coste
Franck
F
Clavaud
Cecile
C
Hetru
Charles
C
Hoffmann
Jules A
JA
Latgé
Jean-Paul
JP
Ferrandon
Dominique
D
Roussel
Alain
A
D000595:Amino Acid Sequence; D000818:Animals; D012831:Bombyx; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray / Q000379:methods; D029721:Drosophila Proteins / Q000737:chemistry* / Q000378:metabolism*; D004331:Drosophila melanogaster; D005656:Fungal Proteins / Q000737:chemistry*; D006458:Hemolymph / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011134:Polysaccharides / Q000737:chemistry; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D047071:beta-Glucans / Q000737:chemistry
D002352:Carrier Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; C516487:GNBP3 protein, Drosophila; D047908:Intracellular Signaling Peptides and Proteins; D008024:Ligands; D011134:Polysaccharides; D047071:beta-Glucans
J Biol Chem. 2004 Jun 18;279(25):26605-11
15084591
Cell. 2006 Dec 29;127(7):1425-37
17190605
J Biol Chem. 1995 Jan 13;270(2):892-7
7822328
Annu Rev Immunol. 2003;21:335-76
12524386
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
EMBO J. 2005 Mar 23;24(6):1277-86
15729357
Nat Rev Microbiol. 2007 Apr;5(4):264-77
17363965
Bioinformatics. 2008 Dec 1;24(23):2780-1
18818215
J Exp Med. 2002 Aug 5;196(3):407-12
12163569
Cold Spring Harb Symp Quant Biol. 1989;54 Pt 1:1-13
2700931
J Microbiol Methods. 2004 Feb;56(2):213-9
14744450
J Biol Chem. 2006 Mar 3;281(9):5771-9
16371356
J Biol Chem. 2000 Feb 18;275(7):4995-5002
10671539
Science. 1999 May 21;284(5418):1313-8
10334979
Nat Rev Immunol. 2007 Nov;7(11):862-74
17948019
J Biol Chem. 2000 Sep 8;275(36):27594-607
10869365
Nature. 2001 Dec 13;414(6865):756-9
11742401
Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9465-9
7568155
Proc Natl Acad Sci U S A. 2007 Apr 17;104(16):6602-7
17409189
EMBO J. 1999 Aug 2;18(15):4087-95
10428948
Protein Sci. 2006 Feb;15(2):304-13
16385003
J Biol Chem. 1988 Aug 25;263(24):12056-62
3136171
Nat Immunol. 2008 Oct;9(10):1165-70
18724373
Nature. 2002 Apr 11;416(6881):640-4
11912488
Nat Immunol. 2004 Nov;5(11):1175-80
15448690
Dev Cell. 2006 Jan;10(1):45-55
16399077
Mol Microbiol. 2008 Jul;69(1):188-200
18466295
Biomacromolecules. 2008 Mar;9(3):783-8
18257529
Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7888-93
8755572
Science. 2003 Dec 19;302(5653):2126-30
14684822
Cell. 2006 Feb 24;124(4):783-801
16497588
Science. 2002 Apr 12;296(5566):298-300
11951031
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
Biochem J. 2004 Sep 15;382(Pt 3):769-81
15214846
Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
12393927
Cell. 1996 Sep 20;86(6):973-83
8808632
J Biol Chem. 2000 Mar 17;275(11):7505-14
10713054
Mol Immunol. 2008 May;45(9):2521-30
18304640
PLoS Biol. 2004 Sep;2(9):E277
15361936
J Mol Biol. 2007 Jan 19;365(3):555-60
17095014
J Biol Chem. 2002 Apr 19;277(16):14281-7
11830593
Cell Mol Life Sci. 2006 Dec;63(23):2710-24
17013558
Science. 2006 Mar 24;311(5768):1761-4
16556841
Biochem Biophys Res Commun. 1986 Dec 30;141(3):1177-84
3028389
Nat Immunol. 2003 Aug;4(8):787-93
12845326
Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13705-10
12359879
Nat Rev Immunol. 2003 May;3(5):371-82
12766759
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Sep 1;65(Pt 9):870-3
19724120
Mol Biol Evol. 2002 May;19(5):748-61
11961108