Identification and characterisation of a dense granule-associated protein in Neospora caninum tachyzoites.

Hemphill, Andrew; Gajendran, N; Sonda, S; Fuchs, N; Gottstein, Bruno; Hentrich, B; Jenkins, M (1998). Identification and characterisation of a dense granule-associated protein in Neospora caninum tachyzoites. International journal for parasitology, 28(3), pp. 429-438. Elsevier

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Neospora caninum is an apicomplexan parasite which is morphologically and ultrastructurally very similar to Toxoplasma gondii. In order to identify molecules involved in host cell entry and subsequent modification of the parasitophorous vacuole, a polyclonal antiserum directed against N. caninum tachyzoites was raised in a rabbit. Subcellular fractionation of tachyzoites was performed using the non-ionic detergent Triton-X-114. Membrane fractions were analysed by immunoblotting using the polyclonal antiserum. One of the immunoreactive protein bands had a mol. wt of 33,000 and was subsequently named Nc-p33. Affinity-purified anti-Nc-p33 antibodies were used to characterise this polypeptide using SDS-PAGE, isoelectric focusing, Western blot analysis and immuno-EM. Nc-p33 was found in two isolates of N. caninum (NC-1 and Liverpool), but could not be detected in T. gondii tachyzoites. Immunogold EM revealed that Nc-p33 constituted a dense granule-associated protein, and Western blotting demonstrated that Nc-p33 was most likely identical to the recently described antigen NCDG1. Shortly after invasion, this dense granule protein was targeted to the parasitophorous vacuole membrane, and, at later timepoints after infection, was also found on the parasitophorous vacuolar network. This suggested that Nc-p33 could play a functional role in the modification of the parasitophorous vacuole and its membrane.

Item Type:	Journal Article (Original Article)
Division/Institute:	05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
UniBE Contributor:	Hemphill, Andrew, Gottstein, Bruno
Subjects:	600 Technology > 630 Agriculture 600 Technology > 610 Medicine & health
ISSN:	0020-7519
Publisher:	Elsevier
Language:	English
Submitter:	Bruno Gottstein
Date Deposited:	23 Jul 2018 09:17
Last Modified:	05 Dec 2022 15:16
PubMed ID:	9559361
URI:	https://boris.unibe.ch/id/eprint/118732