Carbohydrate-binding activity of the type-2 ribosome-inactivating protein SNA-I from elderberry (Sambucus nigra) is a determining factor for its insecticidal activity

2008 ◽  
Vol 69 (17) ◽  
pp. 2972-2978 ◽  
Author(s):  
Shahnaz Shahidi-Noghabi ◽  
Els J.M. Van Damme ◽  
Guy Smagghe
Keyword(s):  
Insecticidal Activity ◽  
Binding Activity ◽  
Carbohydrate Binding ◽  
Ribosome Inactivating Protein ◽  
Sambucus Nigra ◽  
Determining Factor

scholarly journals Mutational analysis of the carbohydrate-binding activity of the NeuAc(α-2,6)Gal/GalNAc-specific type 2 ribosome-inactivating protein from elderberry (Sambucus nigra) fruits

2002 ◽  
Vol 364 (2) ◽  
pp. 587-592 ◽  
Author(s):  
Ying CHEN ◽  
Pierre ROUGE ◽  
Willy J. PEUMANS ◽  
Els J.M. van DAMME
Keyword(s):  
Mutational Analysis ◽  
Binding Activity ◽  
Site Directed Mutagenesis ◽  
Carbohydrate Binding ◽  
Ribosome Inactivating Protein ◽  
Sambucus Nigra ◽  
Original Activity ◽  
Highly Active ◽  
Critical Residue

Sambucus nigra agglutinin I (SNA-I) is a type 2 ribosome-inactivating protein. Site-directed mutagenesis was used to mimic the conversion of the highly active B-chain of fruit-specific SNA (SNA-If) into the completely inactive B-chain of the closely related and naturally occurring loss-of-activity mutant called S. nigra agglutinin lectin-related protein. In the first mutant SNA-If-M1 the high-affinity site 2 of SNA-If was disrupted by replacing the presumed critical residue Asp231 with Glu231. In the double mutant SNA-If-M2, site 1 of SNA-If-M1 was also disrupted by substituting the presumed critical residue Asn48 with Ser48. The parent type 2 ribosome-inactivating protein and both mutants were expressed in Nicotiana tabacum Samsun NN and the recombinant proteins were purified and analysed. Recombinant SNA-If agglutinated rabbit erythrocytes equally well as SNA-If, but both mutants were completely inactive in this test. Binding assays to immobilized galactose and fetuin revealed that the mutation Asp231→Glu231 reduces the affinity of the B-chain for galactose and fetuin by more than 50%. Furthermore, the introduction of the second mutation Asn48→Ser48 reduces the binding activity to less than 20% of the original activity.

scholarly journals Isolation and Molecular Cloning of a Novel Type 2 Ribosome-inactivating Protein with an Inactive B Chain from Elderberry (Sambucus nigra) Bark

1997 ◽  
Vol 272 (13) ◽  
pp. 8353-8360 ◽  
Author(s):  
Els J. M. Van Damme ◽  
Annick Barre ◽  
Pierre Rougé ◽  
Fred Van Leuven ◽  
Willy J. Peumans
Keyword(s):  
Molecular Cloning ◽  
Ribosome Inactivating Protein ◽  
Sambucus Nigra

scholarly journals The major elderberry (Sambucus nigra) fruit protein is a lectin derived from a truncated type 2 ribosome-inactivating protein

1997 ◽  
Vol 12 (6) ◽  
pp. 1251-1260 ◽  
Author(s):  
Els J.M. Damme ◽  
Soma Roy ◽  
Annick Barre ◽  
Pierre Rouge ◽  
Fred Leuven ◽  
...  
Keyword(s):  
Ribosome Inactivating Protein ◽  
Sambucus Nigra

Genomic analysis of C-type lectins

2002 ◽  
Vol 69 ◽  
pp. 59-72 ◽  
Author(s):  
Kurt Drickamer ◽  
Andrew J. Fadden
Keyword(s):  
Biological Effects ◽  
Cell Signalling ◽  
Genomic Analysis ◽  
Binding Activity ◽  
Immunoglobulin Superfamily ◽  
Carbohydrate Binding ◽  
Carbohydrate Recognition ◽  
Calcium Dependent ◽  
Binding Domains ◽  
Carbohydrate Recognition Domains

Many biological effects of complex carbohydrates are mediated by lectins that contain discrete carbohydrate-recognition domains. At least seven structurally distinct families of carbohydrate-recognition domains are found in lectins that are involved in intracellular trafficking, cell adhesion, cell–cell signalling, glycoprotein turnover and innate immunity. Genome-wide analysis of potential carbohydrate-binding domains is now possible. Two classes of intracellular lectins involved in glycoprotein trafficking are present in yeast, model invertebrates and vertebrates, and two other classes are present in vertebrates only. At the cell surface, calcium-dependent (C-type) lectins and galectins are found in model invertebrates and vertebrates, but not in yeast; immunoglobulin superfamily (I-type) lectins are only found in vertebrates. The evolutionary appearance of different classes of sugar-binding protein modules parallels a development towards more complex oligosaccharides that provide increased opportunities for specific recognition phenomena. An overall picture of the lectins present in humans can now be proposed. Based on our knowledge of the structures of several of the C-type carbohydrate-recognition domains, it is possible to suggest ligand-binding activity that may be associated with novel C-type lectin-like domains identified in a systematic screen of the human genome. Further analysis of the sequences of proteins containing these domains can be used as a basis for proposing potential biological functions.

scholarly journals Biochemical and Initial Structural Characterization of the Monocot Chimeric Jacalin OsJAC1

2021 ◽  
Vol 22 (11) ◽  
pp. 5639
Author(s):  
Nikolai Huwa ◽  
Oliver H. Weiergräber ◽  
Christian Kirsch ◽  
Ulrich Schaffrath ◽  
Thomas Classen
Keyword(s):  
Physiological Role ◽  
Basic Function ◽  
Binding Activity ◽  
Carbohydrate Binding ◽  
Lectin Domain ◽  
Reducing Conditions ◽  
Transition Points ◽  
The Individual ◽  
High Yields ◽  
Dirigent Domain

The monocot chimeric jacalin OsJAC1 from Oryza sativa consists of a dirigent and a jacalin-related lectin domain. The corresponding gene is expressed in response to different abiotic and biotic stimuli. However, there is a lack of knowledge about the basic function of the individual domains and their contribution to the physiological role of the entire protein. In this study, we have established a heterologous expression in Escherichia coli with high yields for the full-length protein OsJAC1 as well as its individual domains. Our findings showed that the secondary structure of both domains is dominated by β-strand elements. Under reducing conditions, the native protein displayed clearly visible transition points of thermal unfolding at 59 and 85 °C, which could be attributed to the lectin and the dirigent domain, respectively. Our study identified a single carbohydrate-binding site for each domain with different specificities towards mannose and glucose (jacalin domain), and galactose moieties (dirigent domain), respectively. The recognition of different carbohydrates might explain the ability of OsJAC1 to respond to different abiotic and biotic factors. This is the first report of specific carbohydrate-binding activity of a DIR domain, shedding new light on its function in the context of this monocot chimeric jacalin.

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