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Korean J Parasitol > Volume 32(4):1994 > Article

Original Article
Korean J Parasitol. 1994 Dec;32(4):243-248. English.
Published online Dec 20, 1994.  http://dx.doi.org/10.3347/kjp.1994.32.4.243
Copyright © 1994 by The Korean Society for Parasitology
Identification of surface antigen of Trichomonas vaginalis
D Y Min,*M H Leem,M H Ahn and J S Ryu
Department of Parasitology, College of Medicine, Hanyang University, 133-791 Seoul, Korea.
Received October 27, 1994; Accepted November 11, 1994.

Abstract

Plasma membrane proteins of a Korean isolate of Trichomonas vaginalis HY-1 were fractionated for antigen analysis. Homogenates of T. vaginalis were fractionated by the differential centrifugation using sucrose step-gradient method. The interface layer from the 25%/45% sucrose was collected as a plasma membrane fraction and its purity was examined by transmission electron microscopy. The antigenicity of plasma membrane fraction was analysed by enzyme-linked immunoelectrotransfer blot technique with immune rabbit serum and compared with surface antigen labelled with N-hydroxysuccinimide-biotin. The fluffy fraction of 25%/45% sucrose interface was homogeneous and membrane particles were present as extended sheet and concentric vesicles showing typical trilamellar appearance under transmission electron microscope. Seven fractions at 40, 50, 60, 110, 130, 140 and 150 kDa were identified as the antigenic membrane proteins in EITB with anti HY-1 rabbit serum. The common band at 60 kDa was detected both in antigenic fractions of plasma membrane and surface protein labelled with NHS-biotin. This result indicates that this protein is considered as a major surface antigen of T. vaginalis. The role of this surface antigen at 60 kDa should be studied further.

Figures


Fig. 1
Plasma membrane fraction of T. vaginalis HY-1 isolated by homogenization and sucrose step-gradient method. Membrane particles are present as extended sheet (E) and concentric vesicle (V) (× 10,000).


Fig. 2
Magnification of Fig. 1. (× 30,000).


Fig. 3
Western blot of T. vaginalis HY-1 plasma membrane antigens purified by sucrose step-gradient centrifugation (lane 4) reacted with immune rabbit serum in EITB and immunoprecipitated surface antigen biotinylated prior to lysis (lane 6). Lane 1: High M.W. marker. Lane 2: Amido black stain of TCA precipitated proteins of T. vaginalis HY-1. Lane 3: Immunoblot of whole lysates probed with immune rabbit serum. Lane 5: Biotinylated whole lysate. Lane 7: Immunoprecipitated whole lysate (lysed prior to biotinylation). A: Western blot patterns. B: Schematic diagram of A.

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