title: Search for a dextransucrase minimal motif involved in dextran binding.
abstract: Fourteen truncated forms of Leuconostoc mesenteroides NRRL B512-F dextransucrase, involving N-, C- or N- plus C-terminal domain truncations were tested for their ability to bind dextrans. The shortest fragment (14kDa molecular weight) that still exhibited a strong interaction with dextran was localized between amino acids N1397 and A1527 of the C-terminal domain (GBD-7) and consists of six YG repeats. With a dissociation constant K(d) of 2.8x10(-9)M, this motif shows a very high affinity for isomaltohexaose and longer dextrans, supporting the proposed role of GBD in polymer formation. The potential application of GBD-7 as an affinity tag onto cheap resins like Sephacryl S300HR for rapid purification was evaluated and is discussed.
authors.affiliation: Biological Science Program, Faculty of Science, Chulalongkorn University, 254 Phyathai Road, Patumwan, Bangkok, Thailand.
authors.forename: Surisa
authors.initials: S
authors.lastname: Suwannarangsee
authors.forename: Claire
authors.initials: C
authors.lastname: Moulis
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Potocki-Veronese
authors.forename: Pierre
authors.initials: P
authors.lastname: Monsan
authors.forename: Magali
authors.initials: M
authors.lastname: Remaud-Simeon
authors.forename: Warawut
authors.initials: W
authors.lastname: Chulalaksananukul
mesh_terms: D020816:Amino Acid Motifs; D003911:Dextrans / Q000378:metabolism*; D017353:Gene Deletion; D005936:Glucans / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007934:Leuconostoc / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D016415:Sequence Alignment; D013879:Thioredoxins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism