Isolation and characterization of a cDNA encoding a subtilisin-like serine proteinase (AhSUB) from Acanthamoeba healyi

Hong, Yeon Chul; Kong, Hyun-Hee; Ock, Mee Sun; Kim, In San; Chung, Dong Il

doi:10.1016/s0166-6851(00)00326-1

Cited by 29 publications

(27 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Among them, Kong et al (2000) proposed that a 33 kDa serine proteinase from culture supernatant may play a critical role in pathogenesis of GAE by A. healyi because of the strong activity of the enzyme against main components of extracellular matrix and serum proteins. Hong et al (2000) isolated a cDNA encoding 33 kDa serine proteinase from an A. healyi cDNA library and reported that the proteinase belonged to the subtilisin family. Furthermore, the gene was expressed at higher level in highly-virulent strain (OC-3A) than the low-virulent (KA/E2) or environmental isolate (KA/S8).…”

Section: Discussionmentioning

confidence: 99%

“…In the case of AK, amoeba from environment may attach to the injured surface of cornea and invade the stroma. Up to date, proteinases and adhesion molecules such as laminin binding protein and mannose binding protein have been investigated to understand the pathogenic mechanisms of Acanthamoeba (Hong et al, 2000;Garate et al, 2004;Hong et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

“…The serine protease revealed strikingly potent proteolytic activity against mammalian extracellular matrix proteins including types I and IV collagen and other serum proteins. Hong et al (2000) isolated a cDNA encoding a 33 kDa serine protease (AhSUB), and identified the protease to be a member of subtilase superfamily. Northern blot analysis revealed that AhSUB was expressed at higher levels in the high-virulent strain than low-or avirulent strains.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence

Kim

Kong

et al. 2006

Korean J Parasitol

Self Cite

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence

Kim

Kong

et al. 2006

Korean J Parasitol

Self Cite

View full text Add to dashboard Cite

“…Serine proteinase purified from culture supernatant was reported to play roles in host tissue invasion because of its strong activity against a broad spectrum of extracellular matrix and serum proteins of humans (12). The gene of the purified serine proteinase was characterized and named AhSub based on high homology with bacterial subtilisins (3). Cathepsin L-like cysteine proteinase genes were cloned from Acanthamoeba healyi and Acanthamoeba culbertsoni (3,29).…”

mentioning

confidence: 99%

Intracellular Localization and Trafficking of Serine Proteinase AhSub and Cysteine Proteinase AhCP of Acanthamoeba healyi

et al. 2006

View full text Add to dashboard Cite

Proteinases have been proposed to play important roles in pathogenesis and various biologic actions in Acanthamoeba. Although genetic characteristics of several proteases of Acanthamoeba have been reported, the intracellular localization and trafficking of these enzymes has yet to be studied. In the present study, we analyzed the intracellular localization and trafficking of two proteinases, AhSub and AhCP, of Acanthamoeba healyi by transient transfection. Full-length AhSub-enhanced green fluorescent protein (EGFP) fusion protein was found in intracellular vesicle-like structures of transfected amoebae. Time-lapse photographs confirmed the secretion of the fluorescent material of the vesicle toward the extracellular space. The mutated AhSub, of which the pre or prepro region was deleted, was found to localize diffusely throughout the cytoplasm of the amoeba rather than concentrated in the secretory vesicle. Transfection of the construct containing the pre region only showed the same localization and trafficking of the full-length AhSub. A cysteine proteinase AhCP-EGFP fusion protein showed similar localization in the vesicle-like structure in the amoeba. However, using Lyso Tracker analysis, these vesicular structures of AhCP were confirmed to be lysosomes rather than secretory vesicles. The AhCP construct with a deletion of the prepro region showed a dispersed distribution of fluorescence in the cytoplasm of the cells. These results indicated that AhSub and AhCP would play different roles in Acanthameoba biology and that the pre region of AhSub and pro region of AhCP are important for proper intracellular localization and trafficking of each proteinase.The genus Acanthamoeba, one of the amphizoic amoebae with ubiquitous distribution in human environments, has been known to cause granulomatous amoebic encephalitis in immune-compromised hosts and sight-threatening amebic keratitis, especially in contact lens users (15).Proteinases have been known to play important roles in the metabolism, development, or survival of protozoa (16). Considering the high phagocytic activity of Acanthamoeba as a free-living organism and tissue-invasive behavior as a parasite, proteases would be involved in various processes of Acanthamoeba, such as pathogenesis, nutrient obtainment, and host tissue destruction. To date, several reports on the serine, cysteine, and metalloproteinases of Acanthamoeba were published (9,12,18,19). Serine proteinase purified from culture supernatant was reported to play roles in host tissue invasion because of its strong activity against a broad spectrum of extracellular matrix and serum proteins of humans (12). The gene of the purified serine proteinase was characterized and named AhSub based on high homology with bacterial subtilisins (3). Cathepsin L-like cysteine proteinase genes were cloned from Acanthamoeba healyi and Acanthamoeba culbertsoni (3, 29). However, the specific function of these cysteine proteinases in Acanthamoeba is still unclear. The intracellular localization and trafficking of the e...

show abstract

“…SM; 100 bp ladder, T; trophozoite, C; 3-day cyst, black arrows; reduced genes during encystation, white arrows; increased genes during encystation. (AhSub), which was reported to be a secretory enzyme and a potential virulence factor (Hong et al, 2000). Until now, many proteinases of Acanthamoeba, such as serine, cysteine, and metallo-proteinases, have been suggested to be involved in the pathogenesis or phagocytosis (Mitro et al, 1994;Hong et al, 2000;Kong et al, 2000;Hong et al, 2002;Kim et al, 2006;Serrano-Luna et al, 2006).…”

mentioning

confidence: 99%

Differentially expressed genes of Acanthamoeba castellanii during encystation

Moon

Chung²,

Hong

et al. 2007

Korean J Parasitol

Self Cite

View full text Add to dashboard Cite

Isolation and characterization of a cDNA encoding a subtilisin-like serine proteinase (AhSUB) from Acanthamoeba healyi

Cited by 29 publications

References 21 publications

Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence

Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence

Intracellular Localization and Trafficking of Serine Proteinase AhSub and Cysteine Proteinase AhCP of Acanthamoeba healyi

Differentially expressed genes of Acanthamoeba castellanii during encystation

Contact Info

Product

Resources

About