Isolation, characterization and molecular cloning of the mannose-binding lectins from leaves and roots of garlic (Allium sativum L.).
Two novel lectins were isolated from roots and leaves of garlic. Characterization of the purified proteins indicated that the leaf lectin ASAL is a dimer of two identical subunits of 12 kDa, which closely resembles the leaf lectins from onion, leek and shallot with respect to its molecular structure and agglutination activity. In contrast, the root lectin ASARI, which is a dimer of subunits of 15 kDa, strongly differs from the leaf lectin with respect to its agglutination activity. cDNA cloning of the leaf and root lectins revealed that the deduced amino acid sequences of ASAL and ASARI are virtually identical. Since both lectins have identical N-terminal sequences the larger Mr of the ASARI subunits implies that the root lectin has an extra sequence at its C-terminus. These results not only demonstrate that virtually identical precursor polypeptides are differently processed at their C-terminus in roots and leaves but also indicate that differential processing yields mature lectins with strongly different biological activities. Further screening of the cDNA library for garlic roots also yielded a cDNA clone encoding a protein composed of two tandemly arrayed lectin domains. Since the presumed two-domain root lectin has not been isolated yet, its possible relationship to the previously described two-domain bulb lectin could not be studied at the protein level.
Smeets
K
K
Laboratory for Phytopathology and Plant Protection, Katholieke Universiteit Leuven, Heverlee-Leuven, Belgium.
Van Damme
E J
EJ
Verhaert
P
P
Barre
A
A
Rougé
P
P
Van Leuven
F
F
Peumans
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D003001:Cloning, Molecular; D037222:Collectins; D005737:Garlic / Q000235:genetics* / Q000378:metabolism*; D017343:Genes, Plant; D006384:Hemagglutination; D037102:Lectins / Q000737:chemistry*; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves; D037121:Plant Lectins; D018517:Plant Roots; D010946:Plants, Medicinal; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
D002352:Carrier Proteins; D037222:Collectins; D037102:Lectins; D037121:Plant Lectins; D011994:Recombinant Proteins