title: Characterization of Bacillus thuringiensis Cry toxin binding novel GPI anchored aminopeptidase from fat body of the moth Spodoptera litura.
abstract: Aminopeptidase N (APN) isoforms were identified as candidate receptors for Bacillus thuringiensis Cry toxins from the midgut of several insect species. In this study a partial cDNA encoding aminopeptidase (slfbAPN) was cloned from fat body of the moth Spodoptera litura. In the deduced amino acid sequence the characteristic metallopeptidase sequences, HEXXHX(18)E and GAMENWG were conserved but the sequence showed only 33-39% identity to other insect APNs, which were also reported to be Cry toxin receptors. The presence of a putative GPI anchor signal sequence at the C-terminus indicated that it is a membrane-anchored protein. The slfbAPN expression was restricted to the fat body as suggested by northern blot analysis of different tissues. Biochemical analyses including immunoblotting, ligand blotting and lectin blotting, demonstrated that slfbAPN is a membrane-anchored glycoprotein in the fat body and it binds to Cry toxins.
authors.affiliation: School of Life Sciences, University of Hyderabad, Hyderabad, 500046, India.
authors.forename: Madhusudhan
authors.initials: M
authors.lastname: Budatha
authors.forename: Gargi
authors.initials: G
authors.lastname: Meur
authors.forename: P B
authors.initials: PB
authors.lastname: Kirti
authors.forename: Aparna
authors.initials: A
authors.lastname: Dutta Gupta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis; D001426:Bacterial Proteins; D018826:CD13 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary; D005216:Fat Body / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics; D020539:Sequence Analysis, Protein; D018411:Spodoptera
chemical_list: D001426:Bacterial Proteins; C095416:Cry toxin receptors; D018076:DNA, Complementary; D017261:Glycosylphosphatidylinositols; D019476:Insect Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D018826:CD13 Antigens