Characterization of a new non-toxic two-chain ribosome-inactivating protein and a structurally-related lectin from rhizomes of dwarf elder (Sambucus ebulus L.).
A new N-glycosidase ribosome-inactivating protein (RIP) belonging to the novel family of the nontoxic type 2 RIPs from Sambucaceae has been isolated from rhizomes of dwarf elder (Sambucus ebulus L.) and named ebulin r. Dwarf elder rhizomes also contain a novel monomeric N-Ac-galactosamine-binding lectin that we named SEAII. Ebulin r and SEAII have two isoforms each one, which were readily resolved by ion exchange. Both isoforms of ebulin (ebulins r1 and r2) strongly inhibited protein synthesis in mammalian but not in plant ribosomes by promoting depurination of sensitive ribosomes. Ebulin r and SEAII have apparent molecular masses of 56 and 33.5 kDa, respectively. Ebulins r1 and r2 are composed of two dissimilar subunits (types A-B) of apparent molecular masses of 26 and 30 kDa by disulphide bridges. The rhizome SEAII and the lectins SNA II and SNA III from elder (Sambucus nigra L.) share good amino acid sequence homology. This rhizome ebulin-A chain is more sequence-related to RIP members of cucurbitaceae than to any other plant family. The rhizome ebulin B chain shares a large homology in amino acid sequence with ebulin 1-B chain and SEAII. Anti-ebulin 1 polyclonal antibodies raised in rabbits reacted better with ebulin r1 than with ebulin r2, thus suggesting that both RIP isoforms could have some differences.
Citores
L
L
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Valladolid, Spain.
De Benito
F M
FM
Iglesias
R
R
Ferreras
J M
JM
Argüeso
P
P
Jiménez
P
P
Testera
A
A
Camafeita
E
E
Méndez
E
E
Girbés
T
T
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D010940:Plant Proteins / Q000302:isolation & purification*; D010944:Plants / Q000378:metabolism*; D054788:Ribosome Inactivating Proteins; D054790:Ribosome Inactivating Proteins, Type 2
D010940:Plant Proteins; D054790:Ribosome Inactivating Proteins, Type 2; C107391:ebulin r protein, Sambucus ebulus; D009699:N-Glycosyl Hydrolases; D054788:Ribosome Inactivating Proteins