Search

Jacalin is among the most thoroughly studied lectins. Its carbohydrate-binding site has also been well characterized. It has been postulated that the lower affinity of β-galactosides for jacalin compared with α-galactosides is caused by steric interactions of the substituents in the former with the protein. This issue has been explored energetically and structurally using different appropriate carbohydrate complexes of jacalin. It turns out that the earlier postulation is not correct. The interactions of the substituent with the binding site remain essentially the same irrespective of the anomeric nature of the substitution. This is achieved through a distortion of the sugar ring in β-galactosides. The difference in energy, and therefore in affinity, is caused by a distortion of the sugar ring in β-galactosides. The elucidation of this unprecedented distortion of the ligand as a strategy for modulating affinity is of general interest. The crystal structures also provide a rationale for the relative affinities of the different carbohydrate ligands for jacalin. [ABSTRACT FROM AUTHOR]

Abstract: Compared to puroindolines a and b, grain softness proteins (Gsp1) remained up to now the less characterized members of this wheat specific seed protein family. Due to their low expression levels, their purification remains relatively arduous. We report in the present work on the purification of wheat Gsp, Gsp1b and its post-translational maturation. We showed that Gsp1b underwent different proteolytic cleavages both in the N and C-terminal extremities of the preproprotein. Especially, after the putative signal peptide, a 29 mer peptide is highlighted that is highly truncated in puroindolines (8–9 mer). We also showed that Gsp1b and puroindolines display similar secondary structure but Gsp1b does not interact in vitro with lipids. In addition, by using the iterative threading assembly refinement (I-TASSER) methods, we showed that Gsp1b three-dimensional structure prediction is however identical to both those of puroindolines and of the dicotyledon 2S storage proteins. In contrast with puroindolines, all these data make us question about the role of Gsp1 proteins in endosperm texture and the previously suggested function in plant defense lent to these proteins. [Copyright &y& Elsevier]

Insoluble starch granules stored in plant seeds have generally been considered to be degraded effectively by the combination of amylolytic enzymes following initial attack by de novo synthesized α-amylase at germination. We have shown that rice (Oryza sativa L., var Nipponbare) α-glucosidase isozymes (ONG1, ONG2, and ONG3) are also capable of binding to and degrading starch granules directly, indicating the direct liberation of glucose from starch granules by α-glucosidase at germination. ONG1 and ONG2 are encoded in a distinct locus of the rice genome, while ONG2 and ONG3 are generated by alternative splicing. Interestingly, each of the α-glucosidase isozymes showed different action toward starch granules. In addition, two ONG2 isoforms were found to be produced by post-translational proteolysis. The proteolysis induced changes in binding to and degradation of starch granules. [ABSTRACT FROM AUTHOR]

Abstract: Two isoforms of α-glucosidases (ONG2-I and ONG2-II) were purified from dry rice seeds (Oryza sativa L., var Nipponbare). Both ONG2-I and ONG2-II were the gene products of ONG2 mRNA expressed in ripening seeds. Each enzyme consisted of two components of 6kDa-peptide and 88kDa-peptide encoded by this order in ONG2 cDNA (ong2), and generated by post-translational proteolysis. The 88kDa-peptide of ONG2-II had 10 additional N-terminal amino acids compared with the 88kDa-peptide of ONG2-I. The peptides between 6kDa and 88kDa components (26 amino acids for ONG2-I and 16 for ONG2-II) were removed by post-translational proteolysis. Proteolysis induced changes in adsorption and degradation of insoluble starch granules. We also obtained three α-glucosidase cDNAs (ong1, ong3, and ong4) from ripening seeds. The ONG1, ONG2, and ONG4 genes were situated in distinct locus of rice genome. The transcripts encoding ONG2 and ONG3 were generated by alternative splicing. Members of α-glucosidase multigene family are differentially expressed during ripening and germinating stages in rice. [Copyright &y& Elsevier]