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ESI-MS & NMR based H/D exchange analysis of membrane spanning peptides in model membranes

presented by

Raino K. Hansen
University of Cambridge, UK

February 07, 2003

The Scripps Research Institute, The Committee Lecture Hall


Background:

Raino Hansen studied for his M.Sc. at the Technical University of Denmark and Helsinki University of Technology. He received an open Ph.D. studentship from the Danish Research Academy and decided to study Biochemistry at the University of Cambridge, UK. His Ph.D thesis "Biophysical and biochemical investigation of the M2 peptide of influenza A" was supervised by Dr. R. W. Broadhurst, assistant director of the national NMR facility. The thesis incorporated ESI-MS method development (JASMS, 2002, 13, 1376-1387). Raino Hansen took up a postdoctoral position with Professor N. K. H. Slater in October 2002, with a project to characterise the lack of viral stability of another lipid enveloped virus, herpes simplex virus. Project aims include investigating whether a MALDI-TOF ?fingerprint? of a virus can be correlated with the viral titre measured by lengthy cell culture experiments, an important parameter for the efficiency of gene therapy.

Abstract:

The speaker presents data showing that buffers and detergent can be used in ESI-MS samples, as long as the instrument is carefully operated: Time-resolved hydrogen/deuterium exchange (HDX) studies were performed on the M2 transmembrane peptide of influenza A. Small liposomes were found to be more compatible (J. Fluid. Mech. 378:167) for these HDX/ESI studies than larger ones (PNAS, 97:1389). HDX results demonstrated the importance of lipid membranes in the reconstitution of M2-TM over detergents and solvents. The speaker will also discuss the use of mass spectrometry to evaluate the native like character of model membranes used for structural biology.

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