| Registro completo |
| Provedor de dados: |
ArchiMer
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| País: |
France
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| Título: |
NMR Structure of rALF-Pm3, an Anti-Lipopolysaccharide Factor from Shrimp: Model of the Possible Lipid A-Binding Site
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| Autores: |
Yang, Yinshan
Boze, Helene
Chemardin, Patrick
Padilla, Andre
Moulin, Guy
Tassanakajon, Anchalee
Pugniere, Martine
Roquet, Francoise
Destoumieux Garzon, Delphine
Gueguen, Yannick
Bachere, Evelyne
Aumelas, Andre
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| Data: |
2009-03
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| Ano: |
2009
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| Palavras-chave: |
Ultracentrifugation
Surface plasmon resonance
Septic shock
Structure
NMR
Lipid A
Lipopolysaccharide
Anti lipopolysaccharide factor
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| Resumo: |
The anti-lipopolysaccharide factor ALF-Pm3 is a 98-residue protein identified in hemocytes from the black tiger shrimp Penaeus monodon. It was expressed in Pichia pastoris from the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter as a folded and N-15 uniformly labeled rALF-Pm3 protein. Its 3D structure was established by NMR and consists of three alpha-helices packed against a four-stranded beta-sheet. The C-34-C-55 disulfide bond was shown to be essential for the structure stability. By using surface plasmon resonance, we demonstrated that rALF-Pm3 binds to LPS, lipid A and to OM(R)-174, a soluble analogue of lipid A. Biophysical studies of rALF-Pm3/LPS and rALF-Pm3/OM(R)-174 complexes indicated rather high molecular sized aggregates, which prevented us to experimentally determine by NMR the binding mode of these lipids to rALF-Pm3. However, on the basis of striking structural similarities to the FhuA/LPS complex, we designed an original model of the possible lipid A-binding site of ALF-Pm3. Such a binding site, located on the ALF-Pm3 beta-sheet and involving seven charged residues, is well conserved in ALF-L from Limulus polyphemus and in ALF-T from Tachypleus tridentatus. In addition, our model is in agreement with experiments showing that beta-hairpin synthetic peptides corresponding to ALF-L beta-sheet bind to LPS. Delineating lipid A-binding site of ALFs will help go further in the de novo design of new antibacterial or LPS-neutralizing drugs. (C) 2008 Wiley Periodicals, Inc. Biopolymers 91: 207-220, 2009.
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| Tipo: |
Text
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| Idioma: |
Inglês
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| Identificador: |
http://archimer.ifremer.fr/doc/2009/publication-6320.pdf
DOI:10.1002/bip.21119
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| Editor: |
Wiley / Blackwell
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| Relação: |
http://archimer.ifremer.fr/doc/00000/6320/
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| Formato: |
application/pdf
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| Fonte: |
Biopolymers (0006-3525) (Wiley / Blackwell), 2009-03 , Vol. 91 , N. 3 , P. 207-220
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| Direitos: |
2009 Wiley Periodicals, Inc., A Wiley Company
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