The Membrane-Bound Form of Gene 9 Minor Coat Protein of Bacteriophage M13

eindelijk die kast opruimen!). Summary 108 Samenvatting 111 Curriculum vitae 115 List of Publications 117 Het in dit proefschrift beschreven onderzoek werd mogelijk gemaakt met financiële steun van de Stichting Levenswetenschappen (SLW) van de Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO). Abbreviations ATR-FTIR attenuated total reflection Fourier transform infrared CD circular dichroism C16S g9p with mutation Cys16 to Ser C16S/S32C g9p with mutations Cys16 to Ser and Ser32 to Cys CF 5,6-Carboxyfluorescein DMPC 1,2-dimyristoyl-sn-glycero-3-phosphocholine DMPG 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol DOPC 1,2-dioleoyl-sn-glycero-3-phosphocholine DOPG 1,2-dioleoyl-sn-glycero-3-phosphoglycerol DPPC 1,2-dipalmitoyl-sn-glycero-3-phosphocholine DSC differential scanning calorimetry EDT ethane dithiol Fmoc 9-fluorenylmethoxycarbonyl FTIR Fourier transform infrared G7p, G8p, G9p respectively gene 7-, gene 8-and gene 9-protein HPSEC high performance size exclusion chromatography L/P lipid to protein molar ratio LUV large unilamellar vesicle PMSF phenylmethylsulfonyl fluoride R dichroic ratio RT room temperature SDS sodium dodecyl sulfate TFA trifluoroacetic acid TFE trifluoroethanol 1 Chapter 1 General introduction General Introduction 2 This thesis describes our attempts to gain knowledge about an intriguing protein: the gene 9 minor coat protein (g9p) of bacteriophage M13. Although it is called a 'minor' protein because of the few copies present in the phage particles it may play a major role in initiating the phage assembly process, which occurs in the Escherichia coli membrane. Therefore, it is worth learning about the properties of this small, but important protein. Because g9p is a membrane protein, this introduction starts with a short description of membranes, membrane proteins and protein-lipid interactions. Also several useful methods which are available to reveal properties of this protein in the membrane will be briefly described, with a focus on infrared spectroscopy. Then the introduction continues with the current insights in bacteriophage M13, with special