Pilin C‐terminal peptide binds asialo‐GM1 in liposomes: A 2H‐NMR study

• Published in: Protein science Vol. 6; no. 11; pp. 2459 - 2461
• Main Authors: Jones, David H., Barber, Kathryn R., Grant, Chris W. M., Hodges, Robert S.
• Format: Journal Article
• Language: English
• Published: Bristol Cold Spring Harbor Laboratory Press 01.11.1997
• Subjects: Amino Acid Sequence; Bacterial Outer Membrane Proteins - metabolism; bilayer; Deuterium; deuterium NMR; Fimbriae Proteins; G(M1) Ganglioside - metabolism; glycolipid; Liposomes; Molecular Sequence Data; Nuclear Magnetic Resonance, Biomolecular - methods; peptide; Peptide Fragments - metabolism; Protein Binding; Receptors, Cell Surface - metabolism; spectral symmetry
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1002/pro.5560061120

Wideline 2H‐NMR observations are described demonstrating the interaction of a synthetic peptide (PAK), representing residues 128‐144 of the binding domain of pilin surface protein from Pseudomonas aeruginosa, with a complex glycosphingolipid thought to be its natural receptor. The receptor glycolipid (asialo‐GM1) carried 2H probe nuclei on the terminal and next‐to‐terminal carbohydrate residues and was present as a minor component in fluid phosphatidylcholine liposomes. The peptide induced spectral changes that could be understood as arising from receptor motional changes, without receptor immobilization on the NMR time scale of 104 s−1. Spectral effects were reversed by reduction of the single peptide disulfide bond—a structural feature previously shown to be a determinant of PAK conformation (Campbell AP, McInnes C, Hodges RS, Sykes BD. 1995. Biochemistry 34:6255‐16268). This is the first demonstration of PAK interaction with its epithelial cell receptor in liposomes.