Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthamoeba healyi.
We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healyi OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healyi cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healyi (AhCP1) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues. Cys25, His159, and Asn175. Deduced amino acid sequence analysis indicates that AhCP1 belong to ERFNIN subfamily of C1 peptidases. By Northern blot analysis, no direct correlation was observed between AhCP1 mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that Ahcp1 protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.
Hong
Yeon-Chul
YC
Department of Parasitology, Kyungpook National University School of Medicine, Daegu 700-422, Korea.
Hwang
Mi-Yul
MY
Yun
Ho-Cheol
HC
Yu
Hak-Sun
HS
Kong
Hyun-Hee
HH
Yong
Tai-Soon
TS
Chung
Dong-Il
DI
D000048:Acanthamoeba / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D000562:Amebiasis / Q000469:parasitology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002403:Cathepsins / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004660:Encephalitis / Q000469:parasitology; D015870:Gene Expression; D017125:Genes, Protozoan; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D016415:Sequence Alignment; D014774:Virulence
D016054:DNA, Protozoan; D015800:Protozoan Proteins; D002403:Cathepsins; C455019:AhCP1 enzyme