Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form
As of April 1st VIVO Scientific Profiles will no longer updated for faculty, and the link to VIVO will be removed from the library website. Faculty profile pages will continue to be updated via Interfolio. VIVO will continue being used behind the scenes to update graduate student profiles. Please contact helplib@scripps.edu if you have questions.
How to download citations from VIVO | Alternative profile options

Use of recombinant Entamoeba histolytica cysteine proteinase 1 to identify a potent inhibitor of amebic invasion in a human colonic model

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Melendez-Lopez, S. G.
  • Herdman, S.
  • Hirata, K.
  • Choi, M. H.
  • Choe, Y.
  • Craik, C.
  • Caffrey, C. R.
  • Hansell, E.
  • Chavez-Munguia, B.
  • Chen, Y. T.
  • Roush, William
  • McKerrow, J.
  • Eckmann, L.
  • Guo, J.
  • Stanley, S. L.
  • Reed, S. L.

publication date

  • July 2007

journal

  • Eukaryotic Cell  Journal

abstract

  • Cysteine proteinases are key virulence factors of the protozoan parasite Entamoeba histolytica. We have shown that cysteine proteinases play a central role in tissue invasion and disruption of host defenses by digesting components of the extracellular matrix, immunoglobulins, complement, and cytokines. Analysis of the E. histolytica genome project has revealed more than 40 genes encoding cysteine proteinases. We have focused on E. histolytica cysteine proteinase 1 (EhCP1) because it is one of two cysteine proteinases unique to invasive E. histolytica and is highly expressed and released. Recombinant EhCP1 was expressed in Escherichia coli and refolded to an active enzyme with a pH optimum of 6.0. We used positional-scanning synthetic tetrapeptide combinatorial libraries to map the specificity of the P1 to P4 subsites of the active site cleft. Arginine was strongly preferred at P2, an unusual specificity among clan CA proteinases. A new vinyl sulfone inhibitor, WRR483, was synthesized based on this specificity to target EhCP1. Recombinant EhCP1 cleaved key components of the host immune system, C3, immunoglobulin G, and pro-interleukin-18, in a time- and dose-dependent manner. EhCP1 localized to large cytoplasmic vesicles, distinct from the sites of other proteinases. To gain insight into the role of secreted cysteine proteinases in amebic invasion, we tested the effect of the vinyl sulfone cysteine proteinase inhibitors K11777 and WRR483 on invasion of human colonic xenografts. The resultant dramatic inhibition of invasion by both inhibitors in this human colonic model of amebiasis strongly suggests a significant role of secreted amebic proteinases, such as EhCP1, in the pathogenesis of amebiasis.

subject areas

  • Animals
  • Colon
  • Cysteine Endopeptidases
  • Cysteine Proteinase Inhibitors
  • Disease Models, Animal
  • Entamoeba histolytica
  • Enzyme Activation
  • Humans
  • Hydrogen-Ion Concentration
  • Mice
  • Mice, SCID
  • Protein Conformation
  • Protein Folding
  • Protozoan Proteins
  • Recombinant Proteins
  • Substrate Specificity
  • Sulfones
  • Transplantation, Heterologous
  • Virulence Factors
scroll to property group menus

Identity

PubMed Central ID

  • PMC1951106

International Standard Serial Number (ISSN)

  • 1535-9778

Digital Object Identifier (DOI)

  • 10.1128/ec.00094-07

PubMed ID

  • 17513563
scroll to property group menus

Additional Document Info

start page

  • 1130

end page

  • 1136

volume

  • 6

issue

  • 7

©2022 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support