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


Citation

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

Abstract / Synopsis

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%.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Medicine and Health Science
DOI Number: https://doi.org/10.1107/S2053230X16002016
Publisher: International Union of Crystallography
Keywords: C1ORF123; Hypothetical protein; DUF866; Polycystic ovary syndrome; Bioinformatic analysis
Depositing User: Nabilah Mustapa
Date Deposited: 15 Mar 2018 09:16
Last Modified: 15 Mar 2018 09:16
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1107/S2053230X16002016
URI: http://psasir.upm.edu.my/id/eprint/59649
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