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Title: Structure of Lmaj006129AAA, a hypothetical protein from Leishmania major

Abstract

The crystal structure of a conserved hypothetical protein from L. major, Pfam sequence family PF04543, structural genomics target ID Lmaj006129AAA, has been determined at a resolution of 1.6 Å. The gene product of structural genomics target Lmaj006129 from Leishmania major codes for a 164-residue protein of unknown function. When SeMet expression of the full-length gene product failed, several truncation variants were created with the aid of Ginzu, a domain-prediction method. 11 truncations were selected for expression, purification and crystallization based upon secondary-structure elements and disorder. The structure of one of these variants, Lmaj006129AAH, was solved by multiple-wavelength anomalous diffraction (MAD) using ELVES, an automatic protein crystal structure-determination system. This model was then successfully used as a molecular-replacement probe for the parent full-length target, Lmaj006129AAA. The final structure of Lmaj006129AAA was refined to an R value of 0.185 (R{sub free} = 0.229) at 1.60 Å resolution. Structure and sequence comparisons based on Lmaj006129AAA suggest that proteins belonging to Pfam sequence families PF04543 and PF01878 may share a common ligand-binding motif.

Authors:
;  [1];  [2]; ;  [3];  [4]; ; ;  [3];  [4]; ;  [1];  [2]; ;  [3];  [4];  [3];  [4]; ;  [1] more »;  [2];  [4];  [1];  [2] « less
  1. Department of Biochemistry, University of Washington, Seattle, WA 98195-7742 (United States)
  2. (SGPP) Consortium (United States)
  3. Structural Genomics of Pathogenic Protozoa (SGPP) Consortium (United States)
  4. (United States)
Publication Date:
OSTI Identifier:
22356309
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 3; Other Information: PMCID: PMC2197200; PMID: 16511295; PUBLISHER-ID: tt5005; OAI: oai:pubmedcentral.nih.gov:2197200; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTAL STRUCTURE; CRYSTALLIZATION; DIFFRACTION; FORECASTING; LENGTH; LIGANDS; PROBES; PROTEINS; RESOLUTION; WAVELENGTHS

Citation Formats

Arakaki, Tracy, Le Trong, Isolde, Structural Genomics of Pathogenic Protozoa, Phizicky, Eric, Quartley, Erin, Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, DeTitta, George, Luft, Joseph, Lauricella, Angela, Hauptman-Woodward Institute, Buffalo, NY 14203, Anderson, Lori, Kalyuzhniy, Oleksandr, Structural Genomics of Pathogenic Protozoa, Worthey, Elizabeth, Myler, Peter J., Seattle Biomedical Research Institute, Seattle, WA 98109, Kim, David, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Baker, David, Hol, Wim G. J., Structural Genomics of Pathogenic Protozoa, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Merritt, Ethan A., E-mail: merritt@u.washington.edu, and Structural Genomics of Pathogenic Protozoa. Structure of Lmaj006129AAA, a hypothetical protein from Leishmania major. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106005902.
Arakaki, Tracy, Le Trong, Isolde, Structural Genomics of Pathogenic Protozoa, Phizicky, Eric, Quartley, Erin, Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, DeTitta, George, Luft, Joseph, Lauricella, Angela, Hauptman-Woodward Institute, Buffalo, NY 14203, Anderson, Lori, Kalyuzhniy, Oleksandr, Structural Genomics of Pathogenic Protozoa, Worthey, Elizabeth, Myler, Peter J., Seattle Biomedical Research Institute, Seattle, WA 98109, Kim, David, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Baker, David, Hol, Wim G. J., Structural Genomics of Pathogenic Protozoa, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Merritt, Ethan A., E-mail: merritt@u.washington.edu, & Structural Genomics of Pathogenic Protozoa. Structure of Lmaj006129AAA, a hypothetical protein from Leishmania major. United Kingdom. doi:10.1107/S1744309106005902.
Arakaki, Tracy, Le Trong, Isolde, Structural Genomics of Pathogenic Protozoa, Phizicky, Eric, Quartley, Erin, Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, DeTitta, George, Luft, Joseph, Lauricella, Angela, Hauptman-Woodward Institute, Buffalo, NY 14203, Anderson, Lori, Kalyuzhniy, Oleksandr, Structural Genomics of Pathogenic Protozoa, Worthey, Elizabeth, Myler, Peter J., Seattle Biomedical Research Institute, Seattle, WA 98109, Kim, David, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Baker, David, Hol, Wim G. J., Structural Genomics of Pathogenic Protozoa, Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, Merritt, Ethan A., E-mail: merritt@u.washington.edu, and Structural Genomics of Pathogenic Protozoa. Wed . "Structure of Lmaj006129AAA, a hypothetical protein from Leishmania major". United Kingdom. doi:10.1107/S1744309106005902.
@article{osti_22356309,
title = {Structure of Lmaj006129AAA, a hypothetical protein from Leishmania major},
author = {Arakaki, Tracy and Le Trong, Isolde and Structural Genomics of Pathogenic Protozoa and Phizicky, Eric and Quartley, Erin and Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642 and DeTitta, George and Luft, Joseph and Lauricella, Angela and Hauptman-Woodward Institute, Buffalo, NY 14203 and Anderson, Lori and Kalyuzhniy, Oleksandr and Structural Genomics of Pathogenic Protozoa and Worthey, Elizabeth and Myler, Peter J. and Seattle Biomedical Research Institute, Seattle, WA 98109 and Kim, David and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195 and Baker, David and Hol, Wim G. J. and Structural Genomics of Pathogenic Protozoa and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195 and Merritt, Ethan A., E-mail: merritt@u.washington.edu and Structural Genomics of Pathogenic Protozoa},
abstractNote = {The crystal structure of a conserved hypothetical protein from L. major, Pfam sequence family PF04543, structural genomics target ID Lmaj006129AAA, has been determined at a resolution of 1.6 Å. The gene product of structural genomics target Lmaj006129 from Leishmania major codes for a 164-residue protein of unknown function. When SeMet expression of the full-length gene product failed, several truncation variants were created with the aid of Ginzu, a domain-prediction method. 11 truncations were selected for expression, purification and crystallization based upon secondary-structure elements and disorder. The structure of one of these variants, Lmaj006129AAH, was solved by multiple-wavelength anomalous diffraction (MAD) using ELVES, an automatic protein crystal structure-determination system. This model was then successfully used as a molecular-replacement probe for the parent full-length target, Lmaj006129AAA. The final structure of Lmaj006129AAA was refined to an R value of 0.185 (R{sub free} = 0.229) at 1.60 Å resolution. Structure and sequence comparisons based on Lmaj006129AAA suggest that proteins belonging to Pfam sequence families PF04543 and PF01878 may share a common ligand-binding motif.},
doi = {10.1107/S1744309106005902},
journal = {Acta Crystallographica. Section F},
number = Pt 3,
volume = 62,
place = {United Kingdom},
year = {Wed Mar 01 00:00:00 EST 2006},
month = {Wed Mar 01 00:00:00 EST 2006}
}