Identification and sequence comparison of a cuticular collagen ofBrugia pahangi

Abstract

SUMMARYThe cuticle of filarial nematodes is a specialized extracellular matrix that covers the parasite and protects it from adverse conditions of the environment. As a surface structure it is in direct contact with the host defence mechanisms and therefore plays an important role in the molecular host-parasite relationship. Using polyclonal antisera raised against the insoluble components of the cuticle of the adult filarial parasite Brugia pahangi, we have isolated cDNA clones encoding collagen molecules of the cuticle. The protein domain structure of cDNA clone Bpcol-1 was compared with the known structures of cuticular collagens of the nematodes Brugia malayi, Caenorhabditis elegans, Ascaris suum and Haemonchus contortus, confirming interspecies similarities. Using affinity-purified anti-Bpcol-1 antibodies we identified Bpcol-1 antigenic determinants in different nematode extracts, and determined the localization of such epitopes within the cuticle of B. pahangi.

Collagenase expression by normal human eosinophils

“…The cuticle collagen is similar to human type XII and XIII [34]. As a surface structure it is in direct contact with the host defense mechanisms and possibly plays an important role in the molecular host-parasite relationship [35].…”

Exploring the homolog of a novel proinflammatory microfilarial sheath protein (MfP) of Wuchereria bancrofti in the adult-stage bovine filarial parasite Setaria cervi

“…In brief, parasitic cuticles (outermost layers) were carefully collected by dissecting adult females under a light microscope. Isolated cuticles were chopped into small pieces and somatic materials were removed by treating the cuticle samples through ten subsequent rounds of freezing/thawing, following Bisoffi and Betschart (1996). The cuticle fragments were treated with ethylenediaminetetra-acetic acid (EDTA; 1 m m ), phenylmethyl sulphonyl fluoride (PMSF; 1 m m ), sodium dodecyl sulphate (SDS; 1% v/v), and 2-mercaptoethanol (2-ME; 5% v/v) in phosphate buffered saline (PBS; 50 m m , pH 7.4) for 4 hours at 4°C with constant stirring.…”