TAILIEUCHUNG - Báo cáo khoa học: Expression of the recombinant bacterial outer surface protein A in tobacco chloroplasts leads to thylakoid localization and loss of photosynthesis

Bacterial lipoproteins play crucial roles in host–pathogen interactions and pathogenesis and are important targets for the immune system. A promi-nent example is the outer surface protein A (OspA) ofBorrelia burgdorferi, which has been efficiently used as a vaccine for the prevention of Lyme dis-ease. | ỊFEBS Journal Expression of the recombinant bacterial outer surface protein A in tobacco chloroplasts leads to thylakoid localization and loss of photosynthesis Anna Hennig Katharina Bonfig Thomas Roitsch and Heribert Warzecha University of Wuerzburg Julius-von-Sachs-Institut PharmaceuticalBiology Germany Keywords Lyme disease outer surface protein A plant vaccine protein palmitoylation thylakoid targeting Correspondence H. Warzecha Darmstadt University of Technology Institute of Botany Schnittspahnstrassee 3-5 64287 Darmstadt Germany Fax 49 6151 164630 Tel 49 6151 164024 E-mail warzecha@ Received 7 August 2007 revised 4 September 2007 accepted 5 September 2007 doi Bacterial lipoproteins play crucial roles in host-pathogen interactions and pathogenesis and are important targets for the immune system. A prominent example is the outer surface protein A OspA of Borrelia burgdorferi which has been efficiently used as a vaccine for the prevention of Lyme disease. In a previous study OspA could be produced in tobacco chloroplasts in a lipidated and immunogenic form. To further explore the potential of chloroplasts for the production of bacterial lipoproteins the role of the N-terminal leader sequence was investigated. The amount of recombinant OspA could be increased up to ten-fold by the variation of the insertion site in the chloroplast genome. Analysis of OspA mutants revealed that replacement of the invariant cysteine residue as well as deletion of the leader sequence abolishes palmitolyation of OspA. Also decoration of OspA with an N-terminal eukaryotic lipidation motif does not lead to palmitoyla-tion in chloroplasts. Strikingly the bacterial signal peptide of OspA efficiently targets the protein to thylakoids and causes a mutant phenotype. Plants accumulating OspA at 10 total soluble protein could not grow without exogenously supplied sugars and rapidly died after transfer to soil under greenhouse conditions. The

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