A glycosomal protein (p60) which is predominantly expressed in procyclic Trypanosoma brucei. Characterization and DNA sequence.
Glycosomes are specialized organelles of trypanosomes which contain glycolytic enzymes as their major protein components in Trypanosoma brucei bloodstream form. In the glycosomes of the insect form of T. brucei, additional enzyme activities are found but have not yet been ascribed to a particular protein molecule. In this study, we report the characterization of a 60-kDa glycosomal protein (p60) encoded by a single copy gene which is transcribed into a mRNA of 2.9 kilobases. The gene codes for a protein of 472 amino acids with a molecular mass of 52.5 kDa, suggesting that the mRNA contains large untranslated regions of about 1.4 kilobases. Genomic DNA hybridizations have shown that the gene for p60 is confined to the family of Trypanosomatidae. Sequence comparison confirmed that p60 is not a member of a conserved protein family and does not belong to the group of glycolytic enzymes. p60 is expressed much more strongly in insect form than in bloodstream form trypanosomes. Thus, p60 is the first glycosomal protein observed whose expression is up-regulated during the transition of trypanosomes from the bloodstream to the insect form. The biochemical characterization of p60 demonstrated its capability to bind microtubules and membrane vesicles and to cross-link these structures. These properties might indicate a function in linking glycosomes to the microtubules of the trypanosomal cytoskeleton. However, proteinase K digestion experiments indicate that p60 is not exposed at the outer surface of the glycosomal membrane. The biological role of the microtubule-binding capability of p60 remains unclear, whereas its membrane binding may be of physiological significance inside the glycosome.
Kueng
V
V
Institut für allgemeine Mikrobiologie, Universität Bern, Switzerland.
Schlaeppi
K
K
Schneider
A
A
Seebeck
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003432:Cross-Linking Reagents; D003599:Cytoskeleton / Q000378:metabolism; D008081:Liposomes; D008830:Microbodies / Q000032:analysis* / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D014346:Trypanosoma brucei brucei / Q000032:analysis* / Q000235:genetics / Q000254:growth & development
D003432:Cross-Linking Reagents; D008081:Liposomes; D008869:Microtubule-Associated Proteins