title: Interaction between mosquito-larvicidal Lysinibacillus sphaericus binary toxin components: analysis of complex formation.
abstract: The two components (BinA and BinB) of Lysinibacillus sphaericus binary toxin together are highly toxic to Culex and Anopheles mosquito larvae, and have been employed world-wide to control mosquito borne diseases. Upon binding to the membrane receptor an oligomeric form (BinA2.BinB2) of the binary toxin is expected to play role in pore formation. It is not clear if these two proteins interact in solution as well, in the absence of receptor. The interactions between active forms of BinA and BinB polypeptides were probed in solution using size-exclusion chromatography, pull-down assay, surface plasmon resonance, circular dichroism, and by chemically crosslinking BinA and BinB components. We demonstrate that the two proteins interact weakly with first association and dissociation rate constants of 4.5×10(3) M(-1) s(-1) and 0.8 s(-1), resulting in conformational change, most likely, in toxic BinA protein that could kinetically favor membrane translocation of the active oligomer. The weak interactions between the two toxin components could be stabilized by glutaraldehyde crosslinking. The cross-linked complex, interestingly, showed maximal Culex larvicidal activity (LC50 value of 1.59 ng mL(-1)) reported so far for combination of BinA/BinB components, and thus is an attractive option for development of new bio-pesticides for control of mosquito borne vector diseases.
authors: High Pressure & Synchrotron Radiation Physics Division, Bhabha Atomic Research Centre, Mumbai 400085, India.
authors: Avinash
authors: A
authors: Kale
authors: Ramesh S
authors: RS
authors: Hire
authors: Ashok B
authors: AB
authors: Hadapad
authors: Stanislaus F
authors: SF
authors: D'Souza
authors: Vinay
authors: V
authors: Kumar
mesh_terms: D000818:Animals; D000852:Anopheles / Q000187:drug effects; D001406:Bacillaceae / Q000737:chemistry* / Q000235:genetics; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics / Q000633:toxicity; D003465:Culex / Q000187:drug effects; D007306:Insecticides / Q000737:chemistry* / Q000633:toxicity; D007700:Kinetics
chemical_list: D001427:Bacterial Toxins; D007306:Insecticides; C046134:binB protein, Bacillus sphaericus
keywords: BinA/BinB interactions; Covalent complex; Larvicidal activity; Lysinibacillus sphaericus binary toxin; Mosquito control