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Modulation of endogenous Cysteine Protease Inhibitor (ICP) 1 expression in Entamoeba histolytica affects amoebic adhesion to Extracellular Matrix proteins

 Young Ah Lee  ;  Yumiko Saito-Nakano  ;  Kyeong Ah Kim  ;  Arim Min  ;  Tomoyoshi Nozaki  ;  Myeong Heon Shin 
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Cell Adhesion ; Collagen/metabolism ; Cysteine Proteases/metabolism ; Diamide/pharmacology ; Diazomethane/analogs & derivatives* ; Diazomethane/metabolism ; Dipeptides/genetics ; Dipeptides/metabolism* ; Entamoeba histolytica/enzymology ; Entamoeba histolytica/genetics ; Entamoeba histolytica/metabolism* ; Enzyme Inhibitors/pharmacology ; Ethylmaleimide/pharmacology ; Extracellular Matrix Proteins/metabolism* ; Humans ; Immunoblotting ; Laminin/metabolism ; Sulfhydryl Reagents/pharmacology
Adhesion ; Cysteine protease ; Endogenous cysteine protease inhibitor (ICP) ; Entamoeba histolytica ; Extracellular matrix
Entamoeba histolytica is an enteric tissue-invading protozoan parasite that causes amoebic colitis and occasionally liver abscess in humans. During tissue invasion, amoebic adhesion to host components is an important event for host cell death leading to successful invasion and infection. Among amoebic virulence factors, Gal/GalNAc lectin is known to be major adhesion factor to host cells. In this study, we investigated the role of amoebic secreted CP (Cysteine Proteases) in amoebic adhesion to extracellular matrix (ECM) protein using CP inhibitor and E. histolytica strains in which the endogenous inhibitor of cysteine protease (ICP) 1 gene was overexpressed (ICP1(+)) or repressed by antisense small RNA-mediated gene silencing (ICP1(-)). We found that pretreatment of wild-type amoebae with CP inhibitor E64, or thiol-group modifiers such as diamide and N-Ethylmaleimide resulted in a significant decrease in adhesion to laminin and collagen ECM proteins. Furthermore, ICP1(+) strain, with a reduction of secreted CP activity, exhibited reduced ability by 40% to adhere to laminin. In contrast, ICP1(-) strain, with a 1.9-fold increase of secreted CP activity, showed a two-fold increase in amoebic adherence to laminin compared to the control strain. In addition, total amount of secreted CP5 was decreased in ICP1(+) amoeba. Conversely, total amount of secreted CP1 and mature-form CP5 were increased in ICP1(-) amoeba. We also found that ICP1 was secreted into extracellular milieu. These results suggest that secreted CP activity by E. histolytica may be an important factor affecting adhesion to host proteins, and regulation of CP secretion by ICP plays a major role in pathogenesis. This study provides insight into the CP-mediated tissue pathogenesis in amoeba-invaded lesions during human amoebiasis.
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1. College of Medicine (의과대학) > Dept. of Environmental Medical Biology (환경의생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Kyung Ah(김경아)
Min, A Rim(민아림) ORCID logo https://orcid.org/0000-0003-2938-9630
Shin, Myeong Heon(신명헌) ORCID logo https://orcid.org/0000-0001-8207-6110
Lee, Young Ah(이영아) ORCID logo https://orcid.org/0000-0002-0414-842X
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