Crystallization and preliminary X-ray diffraction study of Lathyrus ochrus isolectin II complexed to the human lactotransferrin N2 fragment.
Isolectin II (LOL II) isolated from the seeds of Lathyrus ochrus has been crystallized in the presence of the N2 fragment (18,500 Da) isolated from human lactotransferrin, which contains an N-acetyllactosamine type biantennary glycan linked to Asn137. This is the first example of a legume lectin crystallized with an N-glycosylprotein. Crystals of the LOL II-N2 complex belong to the tetragonal space group (P4(1)2(1)2 or the enantiomorph) with cell dimensions: a = b = 63.5 A, c = 251.9 A. They diffract well up to at least 3.5 A resolution and more weakly up to 2.8 A resolution. Assuming one functional half-entity in the asymmetric unit, an alpha, beta monomer complexed to one N2 fragment (24,500 Da + 18,500 Da) would give a Vm of 2.95 A3/Da and a solvent content of approximately 58%. SDS/polyacrylamide gels of the dissolved crystals show the presence of both the LOL II and N2 fragment.
Bourne
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Laboratoire de Cristallographie et de Cristallisation des Macromolécules Biologiques, URA no. 1296 du CNRS, Faculté de Médecine Secteur-Nord, Marseille, France.
Nésa
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MP
Rougé
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P
Mazurier
J
J
Legrand
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D
Spik
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Montreuil
J
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Cambillau
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C
D003460:Crystallization; D007887:Fabaceae / Q000737:chemistry*; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry*; D046911:Macromolecular Substances; D037121:Plant Lectins; D010946:Plants, Medicinal; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D014961:X-Ray Diffraction
D037102:Lectins; D046911:Macromolecular Substances; D037121:Plant Lectins; D007781:Lactoferrin