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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
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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. |
format | Online Article Text |
id | pubmed-4498716 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
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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