title: In vivo post-translational modifications of recombinant mussel adhesive protein in insect cells.
abstract: Mussel adhesive proteins (MAPs) have been suggested as promising bioadhesives for diverse application fields, including medical uses. Previously, we successfully constructed and produced a new type of functional recombinant MAP, fp-151, in a prokaryotic Escherichia coli expression system. Even though the E. coli-derived MAP showed several excellent features, such as high production yield and efficient purification, in vitro enzymatic modification is required to convert tyrosine residues to l-3,4-dihydroxyphenyl alanine (dopa) molecules for its adhesive ability, due to the intrinsic inability of E. coli to undergo post-translational modification. In this work, we produced a soluble recombinant MAP in insect Sf9 cells, which are widely used as an effective and convenient eukaryotic expression system for eukaryotic foreign proteins. Importantly, we found that insect-derived MAP contained converted dopa residues by in vivo post-translational modification. In addition, insect-derived MAP also had other post-translational modifications including phosphorylation of serine and hydroxylation of proline that originally occurred in some natural MAPs. To our knowledge, this is the first report on in vivo post-translational modifications of MAP containing dopa and other modified amino acid residues.
authors: Dept. of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors: Seonghye
authors: S
authors: Lim
authors: Kyoung Ro
authors: KR
authors: Kim
authors: Yoo Seong
authors: YS
authors: Choi
authors: Dae-Kyum
authors: DK
authors: Kim
authors: Daehee
authors: D
authors: Hwang
authors: Hyung Joon
authors: HJ
authors: Cha
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D018411:Spodoptera; D053719:Tandem Mass Spectrometry
chemical_list: D017931:DNA Primers; D011506:Proteins; D011994:Recombinant Proteins; C037403:adhesive protein, mussel