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Journal of Bacteriology, May 2009, p. 3220-3225, Vol. 191, No. 10
0021-9193/09/$08.00+0     doi:10.1128/JB.01637-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Crystal Structure of a Chimeric Receptor Binding Protein Constructed from Two Lactococcal Phages ^,

Marina Siponen,¹ Silvia Spinelli,¹ Stéphanie Blangy,¹ Sylvain Moineau,^2,3,4 Christian Cambillau,¹ and Valérie Campanacci^1*

Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS, and Universités Aix-Marseille I & II, Campus de Luminy, Case 932, 13288 Marseille Cedex 09, France,¹ Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire,² Félix d'Hérelle Reference Center for Bacterial Viruses,³ Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Université Laval, Québec City, Québec, Canada G1K 7P4⁴

Received 18 November 2008/ Accepted 5 March 2009

Lactococcus lactis, a gram-positive bacterium widely used by the dairy industry to manufacture cheeses, is subject to infection by a diverse population of virulent phages. We have previously determined the structures of three receptor binding proteins (RBPs) from lactococcal phages TP901-1, p2, and bIL170, each of them having a distinct host range. Virulent phages p2 and bIL170 are classified within the 936 group, while the temperate phage TP901-1 is a member of the genetically distinct P335 polythetic group. These RBPs comprise three domains: the N-terminal domain, binding to the virion particle; a β-helical linker domain; and the C-terminal domain, bearing the receptor binding site used for host recognition. Here, we have designed, expressed, and determined the structure of an RBP chimera in which the N-terminal and linker RBP domains of phage TP901-1 (P335) are fused to the C-terminal RBP domain of phage p2 (936). This chimera exhibits a stable structure that closely resembles the parental structures, while a slight displacement of the linker made RBP domain adaptation efficient. The receptor binding site is structurally indistinguishable from that of native p2 RBP and binds glycerol with excellent affinity.

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* Corresponding author. Mailing address: Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS, and Universités Aix-Marseille I & II, Campus de Luminy, Case 932, 13288 Marseille Cedex 09, France. Phone: 33 491 825 591. Fax: 33 491 266 720. E-mail: valerie.campanacci{at}afmb.univ-mrs.fr

Published ahead of print on 13 March 2009.

Supplemental material for this article is available at http://jb.asm.org/.

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Journal of Bacteriology, May 2009, p. 3220-3225, Vol. 191, No. 10
0021-9193/09/$08.00+0     doi:10.1128/JB.01637-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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