NMR structure of a viral peptide inserted in artificial membranes: a view on the early steps of the birnavirus entry process.
Nonenveloped virus must penetrate the cellular membrane to access the cytoplasm without the benefit of membrane fusion. For birnavirus, one of the peptides present in the virus capsid, pep46 for infectious bursal disease virus, is able to induce pores into membranes as an intermediate step of the birnavirus-penetration pathway. Using osmotic protection experiments, we demonstrate here that pep46 and its pore-forming N-terminal moiety (pep22) form pores of different diameters, 5-8 and 2-4 nm, respectively, showing that both pep46 moieties participate to pore formation. The solution structures of pep46, pep22, and pep24 (the pep46 C-terminal moiety) in different hydrophobic environments and micelles determined by (1)H NMR studies provide structural insights of the pep46 domain interaction. In CDCl(3)/CD(3)OH mixture and in dodecylphosphocholine micelles, the N-terminal domain of pep46 is structured in a long kinked helix, although the C terminus is structured in one or two helices depending upon the solvents used. We also show that the folding and the proline isomerization status of pep46 depend on the type of hydrophobic environment. NMR spectroscopy with labeled phospholipid micelles, differential scanning calorimetry, and plasmon waveguide resonance studies show the peptides lie parallel to the lipid-water interface, perturbing the fatty acid chain packing. All these data lead to a model in which the two domains of pep46 interact with the membrane to form pores.
Galloux
Marie
M
Unité de Pharmacologie Chimique et Génétique, CNRS, UMR 8151, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes, 4 Avenue de l'Observatoire, Paris F-75270 Cedex 06, France.
Libersou
Sonia
S
Alves
Isabel D
ID
Marquant
Rodrigue
R
Salgado
Gilmar F
GF
Rezaei
Human
H
Lepault
Jean
J
Delmas
Bernard
B
Bouaziz
Serge
S
Morellet
Nelly
N
D000818:Animals; D018059:Birnaviridae / Q000378:metabolism*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D004912:Erythrocytes / Q000166:cytology / Q000821:virology; D005227:Fatty Acids / Q000737:chemistry; D006461:Hemolysis; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D012756:Sheep; D012997:Solvents / Q000737:chemistry
D005227:Fatty Acids; D008055:Lipids; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides; D011506:Proteins; D012997:Solvents
Biophys Chem. 1999 Feb 22;76(3):161-83
10074693
Biophys J. 2002 Apr;82(4):1985-94
11916856
Proc Natl Acad Sci U S A. 1991 May 1;88(9):3792-6
1708887
Eur J Biochem. 2000 Apr;267(7):1894-908
10727928
Biopolymers. 1999;51(2):129-44
10397797
Biochemistry. 2003 Jun 3;42(21):6545-58
12767238
J Virol. 1981 Sep;39(3):879-88
6270356
J Biomol NMR. 1998 May;11(4):381-6
9691283
Biophys J. 1997 Sep;73(3):1692-4
19431907
Cell Calcium. 2000 Sep;28(3):137-49
11020376
Biochim Biophys Acta. 1981 Apr 6;642(2):296-312
6269613
Biochem J. 2002 Nov 15;368(Pt 1):171-82
12164787
J Virol. 2002 Oct;76(19):9920-33
12208969
Proteins. 1988;3(4):230-42
2843850
Cell. 2005 Mar 25;120(6):761-72
15797378
Biochem Biophys Res Commun. 1975 Feb 3;62(3):689-95
164186
J Biomol NMR. 2000 Jul;17(3):257-63
10959632
Biochim Biophys Acta. 2002 Aug 31;1564(2):359-64
12175918
Biochemistry. 1998 Aug 25;37(34):11856-63
9718308
Biophys J. 2004 Apr;86(4):2508-16
15041687
EMBO J. 2008 Apr 23;27(8):1289-98
18369316
Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
9757107
J Magn Reson. 2004 May;168(1):153-63
15082261
Biophys J. 2003 Jun;84(6):3751-8
12770881
Proc Natl Acad Sci U S A. 2006 Oct 31;103(44):16496-501
17053074
Protein Sci. 1993 Dec;2(12):2077-84
8298457
Biochem Biophys Res Commun. 1991 Oct 31;180(2):1010-8
1953705
Biophys J. 1997 Aug;73(2):831-8
9251799
Biophys J. 1998 Oct;75(4):1874-85
9746528
EMBO J. 2000 Jan 4;19(1):114-23
10619850
Biochemistry. 1998 Apr 21;37(16):5699-708
9548956
Biophys J. 2001 Sep;81(3):1475-85
11509361
Proc Natl Acad Sci U S A. 1987 Jan;84(1):21-5
3467351
Eur Biophys J. 1999;28(1):48-58
9933923
J Biomol NMR. 2000 Sep;18(1):43-8
11061227
Nature. 1993 Jan 14;361(6408):176-9
8421524
Eur J Biochem. 1997 Sep 15;248(3):938-46
9342250
Biochim Biophys Acta. 2009 Sep;1788(9):1772-81
19427300
J Gen Physiol. 1979 Aug;74(2):157-85
490141
J Biol Chem. 2006 Sep 29;281(39):29278-86
16861222
FEBS Lett. 2002 Dec 4;532(1-2):115-20
12459474
Biochemistry. 1996 Oct 29;35(43):13723-8
8901513
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8245-50
10890923
Biochim Biophys Acta. 1998 Nov 10;1376(3):391-400
9804997
Nature. 1968 Feb 24;217(5130):713-9
5641121
Biochim Biophys Acta. 1991 Mar 18;1063(1):162-70
2015255
FEBS Lett. 1999 Aug 20;457(1):117-21
10486576
Structure. 1994 Apr 15;2(4):271-82
8087554
Biophys J. 2007 Jul 1;93(1):103-12
17416629
Biophys J. 1982 Jan;37(1):353-61
7055627
J Biol Chem. 2007 Jul 13;282(28):20774-84
17488723
Biochim Biophys Acta. 2008 Dec;1778(12):2775-80
18692020
J Virol. 2009 Jul;83(13):6929-33
19369344
J Mol Biol. 2005 Jan 21;345(3):461-74
15581891
Biophys J. 2000 May;78(5):2405-17
10777736
Biochim Biophys Acta. 1999 Mar 4;1417(2):211-23
10082797
J Virol. 2005 Feb;79(4):1992-2000
15681401
Biochimie. 1998 May-Jun;80(5-6):451-9
9782385
J Virol. 1996 Sep;70(9):5832-9
8709201
Biochemistry. 2004 Mar 30;43(12):3590-9
15035629
J Biomol NMR. 1997 Feb;9(2):127-35
9090128
J Biomol NMR. 1992 Nov;2(6):661-5
1490109
Biochim Biophys Acta. 2008 Oct;1778(10):2308-17
18602889
Biochemistry. 2004 Jul 6;43(26):8459-69
15222757
FEBS Lett. 1988 Oct 24;239(1):129-36
3181419
Biochemistry. 2007 Feb 20;46(7):1771-8
17253775
J Virol. 2005 Oct;79(19):12253-63
16160151
Biophys J. 2001 Feb;80(2):801-11
11159447
Phys Rev Lett. 2004 May 14;92(19):198304
15169456
J Virol. 1992 Nov;66(11):6728-34
1404613
Protein Sci. 2004 Aug;13(8):1988-96
15273301
EMBO J. 1996 Apr 15;15(8):1766-77
8617221
Biochim Biophys Acta. 1979 Sep 21;556(2):244-64
534626
Biochim Biophys Acta. 1994 Feb 23;1190(1):181-4
8110813
J Virol. 2002 Mar;76(5):2393-402
11836417
Biochim Biophys Acta. 1999 Dec 15;1462(1-2):141-56
10590306
Methods Enzymol. 2001;339:271-85
11462816
Biochemistry. 2008 May 20;47(20):5565-72
18439024
J Biol Chem. 1994 May 6;269(18):13680-4
8175803
J Virol. 2005 Jun;79(12):7745-55
15919927
Eur J Biochem. 1988 Apr 5;173(1):139-46
3356186
Annu Rev Microbiol. 2002;56:521-38
12142475
Biophys Chem. 2007 Mar;126(1-3):197-200
16730877
Virus Res. 1987 Aug;8(2):127-40
2823498
Chem Biol. 1999 Jul;6(7):473-81
10381406
Biophys J. 2000 Feb;78(2):839-45
10653796
J Virol. 2004 Aug;78(16):8732-45
15280481
Proc Natl Acad Sci U S A. 2008 Apr 1;105(13):5087-92
18375755