Abstract
Protein design is used as an approach to further the understanding of membrane protein assembly, in particular, the assembly of transmembrane (bacterio)chlorophyll-binding pockets. Pigment-protein interaction motifs have been explored by (i) use of model proteins in which the native amino acid sequence in the pigment binding pockets are substantially altered and (ii) theoretical analyses of binding pockets of natural photosynthetic proteins. The bacteriochlorophyll binding sites of light harvesting complex 2, LH2, are replaced by model sites and expressed in vivo by the use of a modified Rhodobacter sphaeroides strain. The artificial helices are shown to bind bacteriochlorophyll and support the assembly of light-harvesting active complexes in the native membrane. A H-bond, which has been introduced at the membrane embedded bacteriochlorophyll/helix model site, is shown to drive the assembly of the model LH2 complex. Statistical analyses of natural (bacterio)chlorophyll binding pockets reveal the presence of distinct interaction motifs at the pigment/helix interface. One example is intra-membrane H-bonding between the pigments and the surrounding polypeptides, particularly between the chlorophylls’ C131 keto carbonyl groups and the residues of the binding helices. With this system at hand, specific interaction motifs, such as the H-bonding motif, and their contribution to the folding and assembly can now be directly addressed within a highly simplified sequence context and in the polypeptides’ native membrane environement.
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Garcia-Martin, A., Kwa, L.G., Jan, M.v., Neil Hunter, C., Braun, P. (2006). Assembly of Model Bacteriochlorophyll Proteins in the Native Lipid Environment. In: Grimm, B., Porra, R.J., Rüdiger, W., Scheer, H. (eds) Chlorophylls and Bacteriochlorophylls. Advances in Photosynthesis and Respiration, vol 25. Springer, Dordrecht. https://doi.org/10.1007/1-4020-4516-6_27
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DOI: https://doi.org/10.1007/1-4020-4516-6_27
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