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Background   Haemonchus contortus , a blood-feeding parasite, is constantly surrounded by large quantities of heme released from the catabolism of host red blood cells. To cope with the toxicity of free heme,  H. contortus  needs to uptake and detoxify the heme, a process believed to be paramount for parasite survival.    Methods  A heme-responsive gene  Hc-hrg-2  was identified which is the homologue of  Ce-hrg-2.  The transcriptional levels in all developmental stages and heme-responsive ability of  Hc-hrg-2  were analyzed by qRT-PCR. Immunofluorescence analysis and cell transfections were performed to analyze the expression pattern of Hc-HGR-2. Statistical analyses were performed with GraghPad Prism 6.0 using Student’s t-test.    Results  To investigate the heme homeostasis of  H. contortus , we first identified a heme-responsive gene  Hc-hrg-2 , a homolog of  Ce-hrg-2  that is involved in heme transport in the hypodermis of  Caenorhabditis elegans . Using qRT-PCR, we showed that  Hc-hrg-2  mRNA was expressed throughout all life-cycle stages of  H. contortus  with the highest level in the third-stage larvae (L3s). Notably, transcription of  Hc-hrg-2  in the exsheathed L3s was significantly upregulated in the presence of high concentration of heme. We found that Hc-HRG-2 protein was mainly located in the hypodermal tissues of adult  H. contortus in vivo  and the endoplasmic reticulum in the transfected mammalian cells. Our  in vitro  assay demonstrated that Hc-HRG-2 is a heme-binding protein with glutathione S-transferase activity and heme had a significant effect on its enzymatic activity when a model substrate 1-chloro-2, 4-dinitrobenzene (CDNB) was used.    Conclusions   Hc-hrg-2  is a heme-responsive gene and engaged in heme homeostasis regulation in hypodermal tissues during the free-living stages of  H. contortus .

Hc-hrg-2, a glutathione transferase gene, regulates heme homeostasis in the blood-feeding parasitic nematode Haemonchus contortus