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Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein

C1ORF123 is a human hypothetical protein found in open reading frame 123 of chromosome 1. The protein belongs to the DUF866 protein family comprising eukaryote-conserved proteins with unknown function. Recent proteomic and bioinformatic analyses identified the presence of C1ORF123 in brain, frontal...

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Main Authors: A. Rahaman, Siti Nurulnabila, Mat Yusop, Jastina, Mohamed Hussein, Zeti Azura, Ho, Kok Lian, Teh, Aik Hong, Waterman, Jitka, Ng, Chyan Leong
Format: Article
Language:English
Published: International Union of Crystallography 2016
Online Access:http://psasir.upm.edu.my/id/eprint/59649/1/Cloning%2C%20expression%2C%20purification%2C%20crystallization%20and%20X-ray%20crystallographic%20analysis%20of%20recombinant%20human%20C1ORF123%20protein.pdf
http://psasir.upm.edu.my/id/eprint/59649/
http://onlinelibrary.wiley.com/doi/10.1107/S2053230X16002016/abstract
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spelling my.upm.eprints.596492018-03-15T01:16:00Z http://psasir.upm.edu.my/id/eprint/59649/ Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein A. Rahaman, Siti Nurulnabila Mat Yusop, Jastina Mohamed Hussein, Zeti Azura Ho, Kok Lian Teh, Aik Hong Waterman, Jitka Ng, Chyan Leong C1ORF123 is a human hypothetical protein found in open reading frame 123 of chromosome 1. The protein belongs to the DUF866 protein family comprising eukaryote-conserved proteins with unknown function. Recent proteomic and bioinformatic analyses identified the presence of C1ORF123 in brain, frontal cortex and synapses, as well as its involvement in endocrine function and polycystic ovary syndrome (PCOS), indicating the importance of its biological role. In order to provide a better understanding of the biological function of the human C1ORF123 protein, the characterization and analysis of recombinant C1ORF123 (rC1ORF123), including overexpression and purification, verification by mass spectrometry and a Western blot using anti-C1ORF123 antibodies, crystallization and X-ray diffraction analysis of the protein crystals, are reported here. The rC1ORF123 protein was crystallized by the hanging-drop vapor-diffusion method with a reservoir solution comprised of 20% PEG 3350, 0.2 M magnesium chloride hexahydrate, 0.1 M sodium citrate pH 6.5. The crystals diffracted to 1.9 Å resolution and belonged to an orthorhombic space group with unit-cell parameters a = 59.32, b = 65.35, c = 95.05 Å. The calculated Matthews coefficient (VM) value of 2.27 Å3 Da−1 suggests that there are two molecules per asymmetric unit, with an estimated solvent content of 45.7%. International Union of Crystallography 2016 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/59649/1/Cloning%2C%20expression%2C%20purification%2C%20crystallization%20and%20X-ray%20crystallographic%20analysis%20of%20recombinant%20human%20C1ORF123%20protein.pdf A. Rahaman, Siti Nurulnabila and Mat Yusop, Jastina and Mohamed Hussein, Zeti Azura and Ho, Kok Lian and Teh, Aik Hong and Waterman, Jitka and Ng, Chyan Leong (2016) Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein. Acta Crystallographica Section F: Structural Biology Communications, 72 (3). pp. 207-213. ISSN 2053-230X http://onlinelibrary.wiley.com/doi/10.1107/S2053230X16002016/abstract 10.1107/S2053230X16002016
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description C1ORF123 is a human hypothetical protein found in open reading frame 123 of chromosome 1. The protein belongs to the DUF866 protein family comprising eukaryote-conserved proteins with unknown function. Recent proteomic and bioinformatic analyses identified the presence of C1ORF123 in brain, frontal cortex and synapses, as well as its involvement in endocrine function and polycystic ovary syndrome (PCOS), indicating the importance of its biological role. In order to provide a better understanding of the biological function of the human C1ORF123 protein, the characterization and analysis of recombinant C1ORF123 (rC1ORF123), including overexpression and purification, verification by mass spectrometry and a Western blot using anti-C1ORF123 antibodies, crystallization and X-ray diffraction analysis of the protein crystals, are reported here. The rC1ORF123 protein was crystallized by the hanging-drop vapor-diffusion method with a reservoir solution comprised of 20% PEG 3350, 0.2 M magnesium chloride hexahydrate, 0.1 M sodium citrate pH 6.5. The crystals diffracted to 1.9 Å resolution and belonged to an orthorhombic space group with unit-cell parameters a = 59.32, b = 65.35, c = 95.05 Å. The calculated Matthews coefficient (VM) value of 2.27 Å3 Da−1 suggests that there are two molecules per asymmetric unit, with an estimated solvent content of 45.7%.
format Article
author A. Rahaman, Siti Nurulnabila
Mat Yusop, Jastina
Mohamed Hussein, Zeti Azura
Ho, Kok Lian
Teh, Aik Hong
Waterman, Jitka
Ng, Chyan Leong
spellingShingle A. Rahaman, Siti Nurulnabila
Mat Yusop, Jastina
Mohamed Hussein, Zeti Azura
Ho, Kok Lian
Teh, Aik Hong
Waterman, Jitka
Ng, Chyan Leong
Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
author_facet A. Rahaman, Siti Nurulnabila
Mat Yusop, Jastina
Mohamed Hussein, Zeti Azura
Ho, Kok Lian
Teh, Aik Hong
Waterman, Jitka
Ng, Chyan Leong
author_sort A. Rahaman, Siti Nurulnabila
title Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
title_short Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
title_full Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
title_fullStr Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
title_full_unstemmed Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein
title_sort cloning, expression, purification, crystallization and x-ray crystallographic analysis of recombinant human c1orf123 protein
publisher International Union of Crystallography
publishDate 2016
url http://psasir.upm.edu.my/id/eprint/59649/1/Cloning%2C%20expression%2C%20purification%2C%20crystallization%20and%20X-ray%20crystallographic%20analysis%20of%20recombinant%20human%20C1ORF123%20protein.pdf
http://psasir.upm.edu.my/id/eprint/59649/
http://onlinelibrary.wiley.com/doi/10.1107/S2053230X16002016/abstract
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score 13.211869

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