Data/hora: 05/06/2024 17:50:48

Provedor de dados: | 5
País: | France
Título: | NMR Structure of rALF-Pm3, an Anti-Lipopolysaccharide Factor from Shrimp: Model of the Possible Lipid A-Binding Site
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.
Data: | 2009-03
Ano: | 2009
Palavras-chave: | Ultracentrifugation; Surface plasmon resonance; Septic shock; Structure; NMR; Lipid A; Lipopolysaccharide; Anti lipopolysaccharide factor.
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.
Tipo: | Text
Idioma: | Inglês

Identificador:

http://archimer.ifremer.fr/doc/2009/publication-6320.pdf

DOI:10.1002/bip.21119

Editor: | Wiley / Blackwell
Relação: | http://archimer.ifremer.fr/doc/00000/6320/
Formato: | application/pdf
Fonte: | Biopolymers (0006-3525) (Wiley / Blackwell), 2009-03 , Vol. 91 , N. 3 , P. 207-220
Fonte: | 2009 Wiley Periodicals, Inc., A Wiley Company