Characterisation and expression of the Fasciola gigantica cathepsin L gene

详细信息    查看全文

• 作者：Yamasaki ; Hiroshi ; Mineki ; Reiko ; Murayama ; Kimie ; Ito ; Akira ; Aoki ; Takashi
• 关键词：Fasciola gigantica ; Cathepsin L ; Gene structure ; Gene family ; Expression ; In vitro processing
• 刊名：International Journal for Parasitology
• 出版年：2002
• 出版时间：July, 2002
• 年：2002
• 卷：32
• 期：8
• 页码：1031-1042
• 全文大小：694 K

文摘

The gene structure of a cathepsin L from Fasciola gigantica was characterised. The gene spans approximately 2.0 kb and comprises four exons and three introns and is a compact gene as in the cases of crustaceous and platyhelminth cathepsins L. Southern blot analysis suggested that a few copies of the genes are sparsely organised in the genome. Of the three intron insertion positions, two of which are in the same position as in the mammalian cathepsin L gene. Phylogenetic analysis revealed that F. gigantica cathepsin L forms a clade with those from Fasciola hepatica, but not with those from Spirometra erinacei and schistosomes. Putative TATA-boxes were found upstream of a transcription initiation site. The sequence analysis of the 5′-upstream of the transcript revealed that the cathepsin L gene is transcribed by cis-splicing fashion. Furthermore, the experiments using recombinant F. gigantica procathepsin L showed that it was processed to an enzymatically active cathepsin L by pH-dependent autocatalysis. However, the pro-peptide deleted cathepsin L showed no enzyme activity, indicating that the pro-region of F. gigantica procathepsin L is essential for the folding and/or refolding of functional cathepsin L. These results are consistent with the observations in mammalian cathepsin L and papain.