Two lectins, called lanceolin and stenodactylin, were purified by affinity chromatography on CL
Sepharose 6B from the caudices of the Passifloraceae Adenia lanceolata and Adenia stenodactyla,
respectively. They are glycoproteins with Mw of 61,243 (lanceolin) and 63,131 daltons
(stenodactylin), consisting of an enzymatic A chain linked to a larger B chain with lectin properties,
with N-terminal amino acid sequences similar to that of volkensin, the toxic lectin from Adenia
volkensii. These two lectins agglutinate red blood cells, inhibit protein synthesis in a cell-free
system as well as in whole cells, and depurinate ribosomes and DNA, but not tRNA or poly(A).
They are highly toxic to cells, in which they induce apoptosis and strongly inhibit protein synthesis,
and to mice, with LD50s 8.16 mg/kg (lanceolin) and 2.76 mg/kg (stenodactylin) at 48 hours after
administration.
Thus, lanceolin and stenodactylin have all the properties of the toxic type 2 ribosomeinactivating
proteins (RIPs).
Further experiments were conducted in order to clarify the effects of these RIPs in cells. We
investigated the cronological relationship between cytotoxic activity, indirectly evaluated as
inhibition of protein synthesis, and loss of cell viability in NB100 cell line.
The induction of apoptosis was assessed by determining caspases 3 and 7 levels, which increase
8-16 hours earlier than the beginning of protein synthesis inhibition. This suggest that the arrest of
protein synthesis is not a central event in the pathway of cell poisoning by RIPs.
The high toxicity and the induction of cell death only by apoptosis and not by necrosis in two
muscular cell lines (TE671 and RD/18) suggest that lanceolin and stenodactylin may be potential
candidates for experimental chemoablation in strabism and blepharospasm.
These results show that lanceolin and stenodactylin are amongst the most potent toxins of plant
origin.
| Identifer | oai:union.ndltd.org:unibo.it/oai:amsdottorato.cib.unibo.it:995 |
| Date | 03 June 2008 |
| Creators | Bortolotti, Massimo <1978> |
| Contributors | Bolognesi, Andrea |
| Publisher | Alma Mater Studiorum - Università di Bologna |
| Source Sets | Università di Bologna |
| Language | Italian |
| Detected Language | English |
| Type | Doctoral Thesis, PeerReviewed |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/restrictedAccess |
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