2y9f

High-resolution Structural Insights on the Sugar-recognition and Fusion Tag Properties of a Versatile b-Trefoil Lectin Domain

spinqualitylabelsall models 2y9f, resolution 1.47Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 2y9f is a 1 chain structure with sequence from Laetiporus sulphureus. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.47Å Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function Q7Z8V1_9APHY Publication Abstract from PubMed In this work, we analyzed at high resolution the sugar-binding mode of the recombinant N-terminal ricin-B domain of the hemolytic protein LSLa (LSL(150)) from the mushroom Laetiporus sulphureus and also provide functional in vitro evidence suggesting that, together with its putative receptor-binding role, this module may also increase the solubility of its membrane pore-forming partner. We first demonstrate that recombinant LSL(150) behaves as an autonomous folding unit and an active lectin. We have determined its crystal structure at 1.47 A resolution and also that of the [LSL(150):(lactose)beta, gamma)] binary complex at 1.67 A resolution. This complex reveals two lactose molecules bound to the beta and gamma sites of LSL(150), respectively. Isothermal titration calorimetry indicates that LSL(150) binds two lactoses in solution with highly different affinities. Also, we test the working hypothesis that LSL(150) exhibits in vivo properties typical of solubility tags. With this aim, we have fused an engineered version of LSL(150) (LSL(t)) to the N-terminal end of various recombinant proteins. All the designed LSL(150)-tagged fusion proteins were successfully produced at high yield, and furthermore, the target proteins were purified by a straightforward affinity procedure on agarose-based matrices due to the excellent properties of LSL(150) as an affinity tag. An optimized protocol for target protein purification was devised, which involved removal of the LSL(150) tag through in-column cleavage of the fusion proteins with His(6)-tagged TEV endoprotease. These results permitted to set up a novel, lectin-based system for production and purification of recombinant proteins in E. coli cells with attractive biotechnological applications. High-resolution structural insights on the sugar-recognition and fusion tag properties of a versatile beta-trefoil lectin domain from the mushroom Laetiporus sulphureus.,Angulo I, Acebron I, de las Rivas B, Munoz R, Rodriguez-Crespo I, Menendez M, Garcia P, Tateno H, Goldstein IJ, Perez-Agote B, Mancheno JM Glycobiology. 2011 Oct;21(10):1349-61. Epub 2011 Jun 1. PMID:21632870 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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