Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143

Acta Crystallogr F Struct Biol Commun. 2015 Jul;71(Pt 7):925-8. doi: 10.1107/S2053230X15009486. Epub 2015 Jun 27.

Abstract

Leishmaniasis is a neglected vector-borne disease with a global prevalence of over 12 million cases and 59,000 annual deaths. Transmission of the parasite requires salivary proteins, including LJL143 from the New World sandfly Lutzomyia longipalpis. LJL143 is a known marker of sandfly exposure in zoonotic hosts. LJL143 was crystallized from soluble protein expressed using Pichia pastoris. X-ray data were collected to 2.6 Å resolution from orthorhombic crystals belonging to space group P2(1)2(1)2(1), with average unit-cell parameters a = 57.39, b = 70.24, c = 79.58 Å. The crystals are predicted to have a monomer in the asymmetric unit, with an estimated solvent content of 48.5%. LJL143 has negligible homology to any reported structures, so the phases could not be determined by molecular replacement. All attempts at S-SAD failed and future studies include experimental phase determination using heavy-atom derivatives.

Keywords: Lutzomyia longipalpis; diagnosis; leishmaniasis; neglected tropical diseases; salivary proteins; sandfly; transmission.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Crystallization
  • Crystallography, X-Ray / methods
  • Gene Expression Regulation
  • Molecular Sequence Data
  • Psychodidae*
  • Salivary Proteins and Peptides / biosynthesis*
  • Salivary Proteins and Peptides / chemistry*
  • Salivary Proteins and Peptides / genetics

Substances

  • Salivary Proteins and Peptides