Dexras1: A human G protein subcloning, expression, and purification optimization and prediction of its structural determinants

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dc.contributor.authorVerma, Ren_US
dc.contributor.authorThapliyal, Aen_US
dc.contributor.authorKumar, N.en_US
dc.date.accessioned2024-11-30T06:00:54Z
dc.date.available2024-11-30T06:00:54Z
dc.date.issued2023-05
dc.description.abstractA small monomeric G protein of the Ras superfamily, Dexras1, involved in cellular signal transduction processes of various pathophysiological processes and interacts with several proteins. Most of these molecular interactions of Dexras1 are attributed to the C-Terminus, but the N-Terminus has not been explored well due to the limited knowledge of its structure and the unavailability of 3D structure Protein Data Bank file in the databases. Our previous studies with in silico homology modeling predicted a structural model of Dexras1 protein and in continuation; the present study was focused on in vitro sub-cloning, expression, and purification of Dexras1 protein to analyze the structural determinants of its N-terminal region. Dexras1 gene was sub-cloned in a customized 6× His tag Maltose Binding Protein vector and expressed in the Escherichia coli BL21 strain. SDS-PAGE analysis suggested that Dexras1 protein was expressed as an insoluble fraction and found the best expression with 1 mM IPTG induction at 37°C for 3–4 h. Then, the expressed protein was purified using Nickel-Nitriloacetic acid resins and analyzed by a circular dichroism (CD) for structural determinants. Spectra obtained by CD outlines preliminary structural information which showed the presence of alpha-helix and beta-sheets in the entire protein, but the N terminus might primarily exist in a non- randomized domain that could have independent functions.en_US
dc.identifier.affiliationsDepartment of Biotechnology, SBAS, Shri Guru Ram Rai University, Dehradun, Uttarakhand, Indiaen_US
dc.identifier.affiliationsDepartment of Life Sciences, Graphic Era (Deemed to be) University, Dehradun, Uttarakhand, Indiaen_US
dc.identifier.affiliationsDepartment of Biotechnology, Graphic Era (Deemed to be) University, Dehradun, Uttarakhand, India.en_US
dc.identifier.citationVerma R, Thapliyal A, Kumar N. . Dexras1: A human G protein subcloning, expression, and purification optimization and prediction of its structural determinants . Journal of Applied Biology & Biotechnology. 2023 May; 11(3): 131-138en_US
dc.identifier.issn2516-7005
dc.identifier.issn2347-212X
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/236580
dc.languageenen_US
dc.publisherOpen Science Publishers LLPen_US
dc.relation.issuenumber3en_US
dc.relation.volume11en_US
dc.source.urihttps://doi.org/10.7324/JABB.2023.59568en_US
dc.subjectDexras1en_US
dc.subjectG proteinen_US
dc.subjectExpressionen_US
dc.subjectPurificationen_US
dc.subjectSub-cloningen_US
dc.subjectSolubility fractions.en_US
dc.titleDexras1: A human G protein subcloning, expression, and purification optimization and prediction of its structural determinantsen_US
dc.typeJournal Articleen_US
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