title: Structural characterization of recombinant human fibroblast growth factor receptor 2b kinase domain upon interaction with omega fatty acids.
abstract: The mutated recombinant kinase domain of human fibroblast growth factor receptor 2b (hFGFR2b) is overexpressed and purified, and its structural changes upon the interaction with three unsaturated fatty acids (UFAs), oleic, linoleic and α-linolenic are studied. This interaction is investigated to find out about the folding and unfolding effect of unsaturated fatty acids on the kinase domain structure of hFGFR2b. Recombinant pLEICS-01 vectors, containing the mutated coding region of hFGFR2b, are expressed in the standard Escherichia coli BL21 (DE3) host cells and purified by Ni2+-NTA affinity chromatography. While polyacrylamide gel electrophoresis characterizes the functionality of recombinant protein, its structural changes are studied in the presence and absence of various concentrations of oleic, α-linolenic and linoleic acids using circular dichroism (CD) and fluorescence spectroscopy. Far ultraviolet CD results show that unsaturated fatty acids do not change the secondary structure of the recombinant kinase domain of hFGFR2b. However, chemical denaturation analysis confirms that all three UFAs destabilize the tertiary structure of recombinant protein. A decrease in the fluorescence intensity without any significant red or blue shift (336±1nm) reflects a variation in the tertiary structure of protein. The direct interaction of the studied UFAs with hFGFR2b reduces the conformational stability of their kinase domains. The structural changes in hFGFR2b in the presence of UFAs may be necessary for hFGFR2b to adjust the signal transduction and regulate the key cellular processes.
authors.affiliation: Department of Biotechnology, School of Para Medicine, Qazvin University of Medical Science, Qazvin, Iran.
authors.forename: Masoumeh
authors.initials: M
authors.lastname: Moghadasi
authors.affiliation: College of Pharmacy, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Dariush
authors.initials: D
authors.lastname: Ilghari
authors.affiliation: Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Sirati-Sabet
authors.affiliation: Centre for Infrastructure Engineering, Western Sydney University, Bld Y, Locked Bag 1797, NSW 2751, Australia; Department of Mechanical Engineering, Australian College of Kuwait, Mishrif, Kuwait City, Kuwait. Electronic address: a.amini@westernsydney.edu.au.
authors.forename: Abbas
authors.initials: A
authors.lastname: Amini
authors.affiliation: Department of Biotechnology, School of Para Medicine, Qazvin University of Medical Science, Qazvin, Iran.
authors.forename: Hamideh
authors.initials: H
authors.lastname: Asghari
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, P.O. Box 34199-15315, Qazvin, Iran. Electronic address: ngheibi@qums.ac.ir.
authors.forename: Nematollah
authors.initials: N
authors.lastname: Gheibi
mesh_terms: D015525:Fatty Acids, Omega-3 / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D011487:Protein Conformation; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D015525:Fatty Acids, Omega-3; D011994:Recombinant Proteins; D051497:Receptor, Fibroblast Growth Factor, Type 2
keywords: Chemical denaturation; Kinase domain; Tertiary structure; Unsaturated fatty acids; hFGFR2b