title: Characterization and identification of the protein partners of Fn3 domain in FnTm2.
abstract: Recently, a novel transmembrane protein was found to be up-regulated in the auditory learning pathway of birds and mammals. The protein, FnTm2, was predicted to have an extracellular fibronectin III (Fn3) domain and a single transmembrane domain. By contrast to other studied Fn3 domains the extracellular domain of FnTm2 bears several cysteine residues, which are predicted to form disulfide bonds. The Fn3 domain of the FnTm2 protein was expressed in DH5-α Escherichia coli (E. coli) cells, purified and characterized by circular dichroism (CD). In order to identify binding partners to Fn3, the isolated protein was incubated with bird brain lysate for a pull down treatment. Of the proteins recognized, myelin basic protein (MBP) was identified as a bona fide partner; it was further characterized for binding to Fn3 in vitro via fluorescence spectroscopy and confirmed via isothermal calorimetry (ITC).
authors: Department of Chemical and Biological Sciences, Polytechnic Institute of NYU, 6 Metrotech Center, Brooklyn, NY 11201, United States.
authors: Peter James
authors: PJ
authors: Baker
authors: Department of Chemical and Biological Sciences, Polytechnic Institute of NYU, 6 Metrotech Center, Brooklyn, NY 11201, United States.
authors: Yan Mei
authors: YM
authors: Chan
authors: Laboratory of Animal Behavior, The Rockefeller University, 1230 York Avenue, New York, NY 10065, United States.
authors: Moritz
authors: M
authors: Hertel
authors: Department of Chemical and Biological Sciences, Polytechnic Institute of NYU, 6 Metrotech Center, Brooklyn, NY 11201, United States; Department of Biochemistry, SUNY Downstate Medical Center, Brooklyn, NY 11203, United States. Electronic address: jmontcla@poly.edu.
authors: Jin Kim
authors: JK
authors: Montclare
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001923:Brain Chemistry; D002151:Calorimetry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry*; D046369:Finches; D008297:Male; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000737:chemistry / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D058849:Protein Refolding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D000199:Actins; D030161:Avian Proteins; D005353:Fibronectins; D004676:Myelin Basic Protein; D011994:Recombinant Proteins