title: Cloning, expression, purification, crystallization and X-ray crystallographic analysis of recombinant human C1ORF123 protein.
abstract: 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 Å(3) Da(-1) suggests that there are two molecules per asymmetric unit, with an estimated solvent content of 45.7%.
authors.affiliation: Institute of Systems Biology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia.
authors.forename: Siti Nurulnabila A
authors.initials: SN
authors.lastname: Rahaman
authors.affiliation: Institute of Systems Biology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia.
authors.forename: Jastina
authors.initials: J
authors.lastname: Mat Yusop
authors.affiliation: Institute of Systems Biology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia.
authors.forename: Zeti-Azura
authors.initials: ZA
authors.lastname: Mohamed-Hussein
authors.affiliation: Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
authors.forename: Kok Lian
authors.initials: KL
authors.lastname: Ho
authors.affiliation: Centre for Chemical Biology, Universiti Sains Malaysia, 11900 Bayan Lepas, Penang, Malaysia.
authors.forename: Aik-Hong
authors.initials: AH
authors.lastname: Teh
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, England.
authors.forename: Jitka
authors.initials: J
authors.lastname: Waterman
authors.affiliation: Institute of Systems Biology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia.
authors.forename: Chyan Leong
authors.initials: CL
authors.lastname: Ng
mesh_terms: D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D006639:Histidine / Q000737:chemistry; D006801:Humans; D026901:Membrane Transport Proteins; D009842:Oligopeptides / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: C000612401:CZIB protein, human; C471213:His-His-His-His-His-His; D026901:Membrane Transport Proteins; D009842:Oligopeptides; D011506:Proteins; D011993:Recombinant Fusion Proteins; D006639:Histidine
keywords: C1ORF123; DUF866; bioinformatic analysis; hypothetical protein; polycystic ovary syndrome