Crystallization and preliminary X-ray diffraction analysis of an anti-H(O) lectin fromLotus tetragonolobusseeds

Abstract

The seed lectin from Lotus tetragonolobus (LTA) has been crystallized. The best crystals grew over several days and were obtained using the vapour-diffusion method at a constant temperature of 293 K. A complete structural data set was collected at 2.00 Å resolution using a synchrotron-radiation source. LTA crystals were found to be monoclinic, belonging to space group P2 1 , with unit-cell parameters a = 68.89, b = 65.83, c = 102.53 Å , = = 90, = 92 . Molecular replacement yielded a solution with a correlation coefficient and R factor of 34.4 and 51.6%, respectively. Preliminary analysis of the molecular-replacement solution indicates a new quaternary association in the LTA structure. Crystallographic refinement is under way.

Temporal changes in the carbohydrates expressed on BG01 human embryonic stem cells during differentiation as embryoid bodies

“…However, AAA (freshwater eel agglutinin) bound neither 12-nor 28-day EBs, suggesting the absence of Fuc α1,3 GlcNAc (glycan array). UEA 1 specifically binds to α1,2-linked fucose ( [4,[21][22][23], glycan array) and the presence of this epitope appears to increase with differentiation, since UEA1 bound to only 15% of undifferentiated cells, but bound to 70 and 80% of 12-and 28-day EBs, respectively. Lotus lectin, which binds most preferentially to terminal α1,2-linked fucosyl groups when a subterminal α1,3 fucose is also present ( [24], glycan array), did not bind to 12-day EBs, but bound readily to 28-day EBs.…”

“…Lotus lectin did not bind to 12-day EBs, yet binds well to the 28-day EBs, indicating the possible appearance of Fuc α1,2 Gal β1,4 (Fuc α1,3) GlcNAc (Lewis y) groups ( [4,[21][22][23][24], glycan array). Subtle differences among lectins of similar nominal specificity could have a large effect on their ability to bind to cells.…”

Identification of a new quaternary association for legume lectins

“…Lotus tetragonolobus agglutinin (LTA) was purchased from Sigma-Aldrich (USA) and was crystallized as described elsewhere (Moreno et al, 2006). Small crystals were submitted to X-ray diffraction at LNLS (Laborató rio Nacional de Luz Síncrotron, Campinas-Brazil) using a synchrotron radiation source (MX1-station).…”

“…Although they encompass different members that are similar in their primary and tertiary structures, several differences have been reported with regard to their mode of quaternary association (Brinda et al, 2005). In general, legume lectins are essentially dimers of dimers, and the different modes of tetramerization are a consequence of a diverse combination of dimeric interfaces seen in these quaternary structures (Moreno et al, 2006;Delatorre et al, 2007). Additionally, the mode of tetramerization of legume lectins is closely related to their affinity for complex oligosaccharides.…”

“…In a previous article, we reported on the crystallization and preliminary X-ray analysis of LTA in the native form (Moreno et al, 2006). In the present study, we have investigated the quaternary structure and oligomerization state of LTA using a combination of Protein Crystallography and small angle X-ray scattering (SAXS) techniques.…”