Crystal structure of the cysteine protease inhibitor 2 from Entamoeba histolytica: functional convergence of a common protein fold.
Cysteine proteases (CP) are key pathogenesis and virulence determinants of protozoan parasites. Entamoeba histolytica contains at least 50 cysteine proteases; however, only three (EhCP1, EhCP2 and EhCP5) are responsible for approximately 90% of the cysteine protease activity in this parasite. CPs are expressed as inactive zymogens. Because the processed proteases are potentially cytotoxic, protozoan parasites have developed mechanisms to regulate their activity. Inhibitors of cysteine proteases (ICP) of the chagasin-like inhibitor family (MEROPS family I42) were recently identified in bacteria and protozoan parasites. E. histolytica contains two ICP-encoding genes of the chagasin-like inhibitor family. EhICP1 localizes to the cytosol, whereas EhICP2 is targeted to phagosomes. Herein, we report two crystal structures of EhICP2. The overall structure of EhICP2 consists of eight β-strands and closely resembles the immunoglobulin fold. A comparison between the two crystal forms of EhICP2 indicates that the conserved BC, DE and FG loops form a flexible wedge that may block the active site of CPs. The positively charged surface of the wedge-forming loops in EhICP2 contrasts with the neutral surface of the wedge-forming loops in chagasin. We postulate that the flexibility and positive charge observed in the DE and FG loops of EhICP2 may be important to facilitate the initial binding of this inhibitor to the battery of CPs present in E. histolytica.
Casados-Vázquez
Luz E
LE
Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Apartado Postal 629, CP 36500, Irapuato, Guanajuato, Mexico.
Lara-González
Samuel
S
Brieba
Luis G
LG
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003461:Crystallography; D018360:Crystallography, X-Ray / Q000379:methods; D057056:Cysteine Proteases / Q000737:chemistry / Q000235:genetics; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry / Q000235:genetics*; D004748:Entamoeba histolytica / Q000201:enzymology / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D015853:Cysteine Proteinase Inhibitors; D009838:Oligodeoxyribonucleotides; D015800:Protozoan Proteins; D057056:Cysteine Proteases; D010447:Peptide Hydrolases