Acetylcholinesterase of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene Identification, Expression, and Biochemical Properties of Recombinant Proteins

Abstract

The southern cattle tick Rhipicephalus (Boophilus) microplus (Bm) is a vector of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. Sequencing and in vitro expression of Bm genes encoding AChE allowed biochemical characterization of three BmAChEs expressed in tick synganglion. rBmAChE1, rBm- AChE2 and rBmAChE3 exhibited substrate preference for acetylthiocholine, high substrate inhibition and sensitivity to AChE-specific inhibitors. OP-insensitivity mutations were demonstrated in rBmAChE1 and rBmAChE3. Gene silencing suggested functional complementation of BmAChEs in vivo. BmAChE genes were amplified in copy number and multiple transcript polymorphisms were expressed in individual tick synganglia for each of the BmAChEs, suggesting allelic diversity within individuals. Studies also identified a gene encoding AChE of the sand fly, Phlebotomus papatasi, a vector of leishmaniasis in humans and animals. Expression of recombinant P. papatasi AChE (rPpAChE) enabled biochemical characterization and identification of effective inhibitors that selectively target rPpAChE.

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Document Details

Document Type
Technical Report
Publication Date
Jan 01, 2013
Accession Number
ADA575351

Entities

People

  • Adalberto A. Pérez De León
  • Alexander P. Tuckow
  • Andrew Y. Li
  • Danett K. Brake
  • Kevin B. Temeyer
  • Pia U. Olafson

Organizations

  • Agricultural Research Service

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Acetylcholinesterases
  • Amino Acids
  • Animal Diseases
  • Animals
  • Cells
  • Chemistry
  • Deoxyribonucleic Acids
  • Genetics
  • Health Services
  • Organophosphates
  • Pest Control
  • Pesticides
  • Proteins
  • Recombinant Proteins
  • Resistance
  • Ribonucleic Acids
  • Ticks

Fields of Study

  • Biology

Readers

  • Molecular Genetics
  • Neurotoxicology
  • Vector-Borne Disease and Entomology