Cloning, molecular analysis and differential cell localisation of the p36 RACK analogue antigen from the parasite protozoon Crithidia fasciculata 1

FEBS Letters

Larraga, Vicente¹  Taladriz, Soraya¹  Gonzalez-Aseguinolaza, Gloria¹  Marquet, Alberto¹ 

[1] Centro de Investigaciones Biologicas, CSIC, Velazquez 144, Madrid 28006, Spain

关键词: p36 RACK analogue;    Protein kinase C activation;    Crithidia fasciculata;    CACK;    Crithidia fasciculata analogue of the receptor for activated protein kinase C (RACK);    DAPI;    4;    6-diamidine-2-phenylindole dihydrochloride;    SSC;    sodium citrate/sodium acetate buffer;    PMSF;    phenylmethylsulphonyl fluoride;    ECL;    enhanced chemiluminescence;   

DOI  :  10.1016/S0014-5793(99)00006-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The family of the RACK molecules (receptors for activated C kinases) are present in all the species studied so far. In the genus Leishmania, these molecules also induce a strong immune reaction against the infection. We have cloned and characterised the gene that encodes the RACK analogue from the parasite trypanosomatid Crithidia fasciculata (CACK). The molecule seems to be encoded by two genes. The sequence analysis of the cloned open reading frame indicates the existence of a high degree of conservation not only with other members of the Trypanosomatidae but also with mammalians. The study of the protein kinase C phosphorylation sites shows the presence of three of them, shared with the mammalian species, additional to those present in the other protozoa suggesting a certain phylogenetic distance between the protozoon Crithidia fasciculata and the rest of the Trypanosomatidae. The CACK-encoded polypeptide shows an additional sequence of four amino acids at the carboxy-terminal end, which produces a different folding of the fragment with the presence of an α-helix instead of the β-sheet usual in all the other species studied. A similar result is elicited at the amino-terminal end by the change of three amino acid residues. The immunolocalisation experiments show that the CACK displays a pattern with a distribution mainly at the plasma membrane, different from that of the related Leishmania species used as control, that displays a distribution close to the nucleus. Altogether, the data suggest that the existence of the structural differences found may have functional consequences.

【 授权许可】

Unknown