PURIFICATION AND CHARACTERIZATION OF A SECRETORY SERINE PROTEINASE OF ACANTHAMOEBA HEALYI ISOLATED FROM GAE

We purified and characterized a serine proteinase secreted by Acanthamoeba healyi to evaluate it as a possible virulence factor in the pathogenesis of granulomatous amoebic encephalitis (GAE). Ammonium sulfate precipitated culture supernatant of A. healyi OC-3A strain was purified by chromatography on CM-Sepharose, Sephacryl-S200, and Q-2 anion-exchange columns. The purified 33-kDa enzyme had a pH optimum of 8.0 and a temperature optimum of 40 C. Phenylmethylsulfonylfluoride and diisopropyl fluorophosphate, serine proteinase inhibitors, diminished activity of the enzyme to near zero. In addition to types I and IV collagen and fibronectin, the main components of the extracellular matrix, other proteins such as fibrinogen, IgG, IgA, albumin, and hemoglobin were also degraded by the enzyme. The broad substrate specificity of this secreted serine proteinase suggests that it may play an important role in pathogenesis of GAE by A. healyi.

[1]  D. Jefferson,et al.  Role of Carbohydrate-mediated Adherence in Cytopathogenic Mechanisms of Acanthamoeba * , 1998, The Journal of Biological Chemistry.

[2]  D. Toney,et al.  Resistance of Acanthamoeba species to complement lysis. , 1998, Journal of Parasitology.

[3]  J. Niederkorn,et al.  Mannose Induces the Release of Cytopathic Factors from Acanthamoeba castellanii , 1998, Infection and Immunity.

[4]  Y. Chung,et al.  Cysteine protease activities during maturation stages of Paragonimus westermani. , 1997, The Journal of parasitology.

[5]  Z. Fishelson,et al.  Schistosoma mansoni: evidence for a 28-kDa membrane-anchored protease on schistosomula. , 1996, Experimental parasitology.

[6]  R. Gerard,et al.  Characterization of a plasminogen activator produced by Acanthamoeba castellanii. , 1995, Molecular and biochemical parasitology.

[7]  C. Kemper,et al.  Disseminated acanthamoeba infection in patients with AIDS: case reports and review. , 1995, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[8]  J. McKerrow,et al.  Partial characterization of the proteolytic secretions of Acanthamoeba polyphaga. , 1994, Experimental parasitology.

[9]  H. Moura,et al.  Acanthamoeba healyi N. Sp. and the Isoenzyme and Immunoblot Profiles of Acanthamoeba spp., Groups 1 and 3 , 1992, The Journal of protozoology.

[10]  Z. Fishelson,et al.  Recent Insights into the Structure and Function of a Larval Proteinase Involved in Host Infection by a Multicellular Parasite , 1991, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[11]  J. Jonckheere.  Ecology of Acanthamoeba , 1991 .

[12]  R. Kowalski,et al.  Growth of acanthamoeba on human corneal epithelial cells and keratocytes in vitro. , 1991, Investigative ophthalmology & visual science.

[13]  J. McCulley,et al.  In vivo and in vitro collagenolytic activity of Acanthamoeba castellanii. , 1990, Investigative ophthalmology & visual science.

[14]  A. Barrett,et al.  Affinity purification and biochemical characterization of histolysin, the major cysteine proteinase of Entamoeba histolytica. , 1988, The Biochemical journal.

[15]  J. McKerrow,et al.  The major neutral proteinase of Entamoeba histolytica , 1986, The Journal of experimental medicine.

[16]  B. Na,et al.  Cloning and Expression of a Serine Proteinase Gene Fragment from Acanthamoeba culbertsoni , 1998 .

[17]  C. Song,et al.  Purification and characterization of a serine proteinase from Acanthamoeba culbertsoni , 1996 .

[18]  J. Hassell,et al.  Extracellular matrix : a practical approach , 1995 .

[19]  P. Rosenthal,et al.  The proteases and pathogenicity of parasitic protozoa. , 1993, Annual review of microbiology.

[20]  J. McCulley,et al.  Characterization and pathogenic potential of a soil isolate and an ocular isolate of Acanthamoeba castellanii in relation to Acanthamoeba keratitis. , 1992, Current eye research.

[21]  J. D. de Jonckheere.  Ecology of Acanthamoeba. , 1991, Reviews of infectious diseases.