Small-angle neutron scattering studies suggest the mechanism of BinAB protein internalization.

ABSTRACT: Small-angle neutron scattering (SANS) is one of the most widely used neutron-based approaches to study the solution structure of biological macromolecular systems. The selective deuterium labelling of different protein components of a complex provides a means to probe conformational changes in multiprotein complexes. The Lysinibacillus sphaericus mosquito-larvicidal BinAB proteins exert toxicity through interaction with the receptor Cqm1 protein; however, the nature of the complex is not known. Rationally engineered deuterated BinB (dBinB) protein from the L. sphaericus ISPC-8 species was synthesized using an Escherichia coli-based protein-expression system in M9 medium in D2O for 'contrast-matched' SANS experiments. SANS data were independently analysed by ab initio indirect Fourier transform-based modelling and using crystal structures. These studies confirm the dimeric status of Cqm1 in 100% D2O with a longest intramolecular vector (D max) of ?94?Å and a radius of gyration (R g) of ?31?Å. Notably, BinB binds to Cqm1, forming a heterodimeric complex (D max of ?129?Å and R g of ?40?Å) and alters its oligomeric status from a dimer to a monomer, as confirmed by matched-out Cqm1-dBinB (D max of ?70?Å and R g of ?22?Å). The present study thus provides the first insight into the events involved in the internalization of larvicidal proteins, likely by raft-dependent endocytosis.

SUBMITTER: Sharma M 

PROVIDER: S-EPMC7055391 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature