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

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 zo...

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Autores principales: Kelleher, Alan, Liu, Zhuyun, Seid, Christopher A., Zhan, Bin, Asojo, Oluwatoyin A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498716/
https://www.ncbi.nlm.nih.gov/pubmed/26144240
http://dx.doi.org/10.1107/S2053230X15009486
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author Kelleher, Alan
Liu, Zhuyun
Seid, Christopher A.
Zhan, Bin
Asojo, Oluwatoyin A.
author_facet Kelleher, Alan
Liu, Zhuyun
Seid, Christopher A.
Zhan, Bin
Asojo, Oluwatoyin A.
author_sort Kelleher, Alan
collection PubMed
description 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.
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spelling pubmed-44987162015-07-28 Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143 Kelleher, Alan Liu, Zhuyun Seid, Christopher A. Zhan, Bin Asojo, Oluwatoyin A. Acta Crystallogr F Struct Biol Commun Research Communications 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. International Union of Crystallography 2015-06-27 /pmc/articles/PMC4498716/ /pubmed/26144240 http://dx.doi.org/10.1107/S2053230X15009486 Text en © Kelleher et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Communications
Kelleher, Alan
Liu, Zhuyun
Seid, Christopher A.
Zhan, Bin
Asojo, Oluwatoyin A.
Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title_full Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title_fullStr Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title_full_unstemmed Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title_short Expression, purification, crystallization and crystallographic study of Lutzomyia longipalpis LJL143
title_sort expression, purification, crystallization and crystallographic study of lutzomyia longipalpis ljl143
topic Research Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498716/
https://www.ncbi.nlm.nih.gov/pubmed/26144240
http://dx.doi.org/10.1107/S2053230X15009486
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