Fig. 5
Identification of T. solium metacestode (TsM) cellular proteins bound to calcareous corpuscle (CC)-fasciclin (TsMFas1 or TsMFas2) binary complex. a Immunoblot analysis of scolex/neck (SN) proteins depleted of TsMFas1/2 proteins. SN proteins were incubated with protein G-coupled anti-rTsMFas1/2 antibodies, and unbound proteins were eluted in flow-through fractions. Proteins (10 μg) were separated by 8% SDS-PAGE under reducing conditions and transblotted to a nitrocellulose membrane. Blots were probed with anti-rTsMFas1 or anti-rTsMFas2 antibody (1:2000 dilution). Immunoreactive signals were developed using ECL after 2 min exposure. Lane SN: whole SN proteins; Lane SNFas1/2-: SN protein depleted of Fas1/2. Abbreviation: M r., molecular weight in kDa. b TsM SN proteins depleted of Fas1 and Fas2 proteins (10 μg) were incubated with CC-rTsMFas1 or CC-rTsMFas2 binary complex (each 10 μl) and precipitated by centrifugation. Pellets were resuspended in 2× SDS-PAGE reducing sample buffer, separated on 15% gels and stained with CBB. Lane SN: scolex/neck protein (10 μg); Lane SNFas1/2-: SN protein depleted of Fas1/2 (10 μg); Lane CC: purified calcareous corpuscle (10 μl) only; Lane CC + rFas1 or 2: purified CC was incubated with rFas1 or rFas2 protein; Lane CC + SNFas1/2-: Fas1/2 depleted SN proteins were incubated with CC; Lane CC + rFas1/2 + SNFas1/2-: CC-rFas1 or CC-rFas2 binary complex was incubated with Fas1 and Fas2 depleted SN proteins. Abbreviation: M r., molecular weight in kDa. c Identification of protein repertoire for CC-Fas complex. Each protein band (7–14 and A-C) was processed for protein identification by LC-ESI-MS/MS. Independent duplicated biological samples were analysed. d Construction of protein-protein interaction network mediated by CC-Fas1 or CC-Fas2 complex by STRING algorithm ver10.0 (http://string-db.org/). Correlated interactions extracted from the platyhelminth proteins are presented with their predicted functional partners