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Flexibility of truncated and full-length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

Pijning, T., Vujicic - Zagar, A., Kralj, S., Dijkhuizen, L. & Dijkstra, B. W., May-2014, In : Febs Journal. 281, 9, p. 2159-2171 13 p.

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  • Flexibility of truncated and full-length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

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DOI

Glucansucrase enzymes synthesize high-molecular-mass extracellular -glucan polysaccharides from sucrose. Previously, the crystal structure of truncated glucansucrase glucosyltransferase (GTF)180-N from Lactobacillusreuteri 180 (lacking the N-terminal domain) revealed an elongated overall structure with two remote domains (IV and V) extending away from the core. By contrast, a new crystal form of the -1,6/-1,3 specific glucansucrase GTF180-N shows an approximate 120(o) rotation of domain V about a hinge located between domains IV and V, giving a much more compact structure than before. Positional variability of domain V in solution is confirmed by small angle X-ray scattering experiments and rigid-body ensemble calculations. In addition, small angle X-ray scattering measurements of full-length GTF180 also provide the first structural data for a full-length glucansucrase, showing that the enzyme has an almost symmetric boomerang-like molecular shape, with a bend likely located between domains IV and V. The similar to 700-residue N-terminal domain, which is not present in the crystal structures, extends away from domain V and the catalytic core of the enzyme. We conclude that, as a result of the hinge region, in solution, GTF180-N (and likely also the full-length GTF180) shows conformational flexibility; this may be a general feature of GH70 glucansucrases.

Database center dot Structural data for GTF180-N II have been deposited in the Protein Data Bank under accession code .

Original languageEnglish
Pages (from-to)2159-2171
Number of pages13
JournalFebs Journal
Volume281
Issue number9
Publication statusPublished - May-2014

    Keywords

  • crystal structure, glucansucrase, GTF180, rigid-body ensemble calculations, SAXS, SMALL-ANGLE SCATTERING, X-RAY-SCATTERING, STRUCTURAL-CHARACTERIZATION, PROTEINS, MECHANISM, REVEALS, GENES

ID: 15903594