Characterization of multisugar-binding C-type lectin (SpliLec) from a bacterial-challenged cotton leafworm, Spodoptera littoralis.
BACKGROUND
Various proteins that display carbohydrate-binding activity in a Ca(2+)-dependent manner are classified into the C-type lectin family. They have one or two C-type carbohydrate-recognition domains (CRDs) composed of 110-130 amino acid residues in common. C-type lectins mediate cell adhesion, non-self recognition, and immuno-protection processes in immune responses and thus play significant roles in clearance of invaders, either as cell surface receptors for microbial carbohydrates or as soluble proteins existing in tissue fluids. The lectin of Spodoptera littoralis is still uncharacterized.


METHODS
A single orf encoding a deduced polypeptide consisting of an 18-residue signal peptide and a 291-residue mature peptide, termed SpliLec, was isolated from the haemolymph of the cotton leafworm, S. littoralis, after bacterial challenge using RACE-PCR. Sequence analyses of the data revealed that SpliLec consists of two CRDs. Short-form CRD(1) and long-form CRD(2) are stabilized by two and three highly conserved disulfide bonds, respectively. SpliLec shares homology with some dipteran lectins suggesting possible common ancestor. The purified SpliLec exhibited a 140-kDa molecular mass with a subunit molecular mass of 35 kDa. The hemagglutination assays of the SpliLec confirmed a thermally stable, multisugar-binding C-type lectin that binds different erythrocytes. The purified SpliLec agglutinated microorganisms and exhibited comparable antimicrobial activity against gram (+) and gram (-) bacteria too.


CONCLUSIONS
Our results suggested an important role of the SpliLec gene in cell adhesion and non-self recognition. It may cooperate with other AMPs in clearance of invaders of Spodoptera littoralis.
Department of Entomology, Faculty of Science, Cairo University, Giza, Egypt. alaaseufi@yahoo.com
AlaaEddeen M
AM
Seufi
Fatma H
FH
Galal
Elsayed E
EE
Hafez
D000595:Amino Acid Sequence; D000818:Animals; D018407:Bacterial Physiological Phenomena; D001483:Base Sequence; D050260:Carbohydrate Metabolism; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D006386:Hemagglutination Tests; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D018411:Spodoptera / Q000378:metabolism* / Q000382:microbiology
D037181:Lectins, C-Type; D004247:DNA