Identification of a 44 kDa protein localized within the endoplasmic reticulum of Trypanosoma brucei brucei.

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  • Author(s): Nandan D;Nandan D; Wells CW; Ndegwa D; Pearson TW
  • Source:
    Parasitology [Parasitology] 1995 Sep; Vol. 111 ( Pt 3), pp. 313-23.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Cambridge University Press Country of Publication: England NLM ID: 0401121 Publication Model: Print Cited Medium: Print ISSN: 0031-1820 (Print) Linking ISSN: 00311820 NLM ISO Abbreviation: Parasitology Subsets: MEDLINE
    • Publication Information:
      Original Publication: London, New York, Cambridge University Press.
    • Subject Terms:
      Protozoan Proteins/*metabolism ; Trypanosoma brucei brucei/*metabolism; Amino Acid Sequence ; Animals ; Antibodies, Protozoan ; Endoplasmic Reticulum/genetics ; Endoplasmic Reticulum/metabolism ; Microscopy, Immunoelectron ; Molecular Chaperones/chemistry ; Molecular Chaperones/metabolism ; Molecular Sequence Data ; Molecular Weight ; Pronase ; Protozoan Proteins/chemistry ; Protozoan Proteins/genetics ; Rats ; Ruthenium Red ; Trypanosoma brucei brucei/genetics ; Trypanosoma brucei brucei/ultrastructure
    • Abstract:
      Immunoaffinity chromatography and gel electrophoresis were used to isolate a 44 kDa protein that was bound to a 72 kDa chaperone in Trypanosoma brucei brucei. A polyclonal antiserum to the 44 kDa protein was raised in rats and employed in conjunction with chromatography using DEAE-cellulose, Sephacryl S-300, and hydroxyapatite to purify the protein from membranes of bloodstream forms of the trypanosomes. Immunoblot analysis using this antiserum revealed a protein doublet of 44/45 kDa in T. b. brucei and a single protein band of 53 kDa in almost equivalent amounts throughout the life-cycle stages of T. congolense. Indirect immunofluorescence using affinity-purified antibodies specific for the 44 kDa protein showed labelling of the perinuclear area and reticular system extending throughout the parasites, suggesting that this protein was located in the endoplasmic reticulum. Localization of the 44 kDa molecule in the endoplasmic reticulum was confirmed by immunoelectron microscopy. Protease protection experiments demonstrated that the epitopes bound by antibody were buried within the membrane or towards the lumenal face of the endoplasmic reticulum. Ruthenium Red overlay of nitrocellulose blots containing the 44/45 kDa doublet suggested that the molecules have the potential to bind calcium. The N-terminal amino acid sequence of the 44 kDa protein showed no sequence similarity to any proteins in the database.
    • Accession Number:
      0 (Antibodies, Protozoan)
      0 (Molecular Chaperones)
      0 (Protozoan Proteins)
      11103-72-3 (Ruthenium Red)
      EC 3.4.24.- (Pronase)
    • Publication Date:
      Date Created: 19950901 Date Completed: 19951113 Latest Revision: 20190818
    • Publication Date:
      20240829
    • Accession Number:
      10.1017/s0031182000081865
    • Accession Number:
      7567099