title: Disruption of a specific molecular interaction with a bound lipid affects the thermal stability of the purple bacterial reaction centre.
abstract: Relatively little is known about the functions of specific molecular interactions between membrane proteins and membrane lipids. The structural and functional consequences of disrupting a previously identified interaction between a molecule of the diacidic lipid cardiolipin and the purple bacterial reaction centre were examined. Mutagenesis of a highly conserved arginine (M267) that is responsible for binding the head-group of the cardiolipin (to leucine) did not affect the rate of photosynthetic growth, the functional properties of the reaction centre, or the X-ray crystal structure of the complex (determined to a resolution of 2.8 A). However, the thermal stability of the protein was compromised by this mutation, part of the reaction centre population showing an approximately 5 degrees C decrease in melting temperature in response to the arginine to leucine mutation. The crystallised mutant reaction centre also no longer bound detectable amounts of cardiolipin at this site. Taken together, these observations suggest that this particular protein-lipid interaction contributes to the thermal stability of the complex, at least when in detergent micelles. These findings are discussed in the light of proposals concerning the unfolding processes that occur when membrane proteins are heated, and we propose that one function of the cardiolipin is to stabilise the interaction between adjacent membrane-spanning alpha-helices in a region where there are no direct protein-protein interactions.
authors.affiliation: Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, BS8 1TD Bristol, UK.
authors.forename: Paul K
authors.initials: PK
authors.lastname: Fyfe
authors.forename: Neil W
authors.initials: NW
authors.lastname: Isaacs
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Cogdell
authors.forename: Michael R
authors.initials: MR
authors.lastname: Jones
mesh_terms: D002152:Calorimetry, Differential Scanning; D002308:Cardiolipins / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D006358:Hot Temperature; D008563:Membrane Lipids / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011489:Protein Denaturation; D020560:Proteobacteria / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002308:Cardiolipins; D008563:Membrane Lipids; D045322:Photosynthetic Reaction Center Complex Proteins