Redox Capacity of an Extracellular Matrix Protein Associated with Adhesion in Mytilus californianus

Biochemistry. 2016 Apr 5;55(13):2022-30. doi: 10.1021/acs.biochem.6b00044. Epub 2016 Mar 21.

Abstract

Adhesive mussel foot proteins (Mfps) rely in part on DOPA (3,4-dihydroxyphenyl-l-alanine) side chains to mediate attachment to mineral surfaces underwater. Oxidation of DOPA to Dopaquinone (Q) effectively abolishes the adsorption of Mfps to these surfaces. The thiol-rich mussel foot protein-6 (Mfp-6) rescues adhesion compromised by adventitious DOPA oxidation by reducing Q back to DOPA. The redox chemistry and kinetics of foot-extracted Mfp-6 were investigated by using a nonspecific chromogenic probe to equilibrate with the redox pool. Foot-extracted Mfp-6 has a reducing capacity of ~17 e(-) per protein; half of this comes from the cysteine residues, whereas the other half comes from other constituents, probably a cohort of four or five nonadhesive, redox-active DOPA residues in Mfp-6 with an anodic peak potential ~500 mV lower than that for oxidation of cysteine to cystine. At higher pH, DOPA redox reversibility is lost possibly due to Q scavenging by Cys thiolates. Analysis by one- and two-dimensional proton nuclear magnetic resonance identified a pronounced β-sheet structure with a hydrophobic core in foot-extracted Mfp-6 protein. The structure endows redox-active side chains in Mfp-6, i.e., cysteine and DOPA, with significant reducing power over a broad pH range, and this power is measurably diminished in recombinant Mfp-6.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adhesiveness
  • Animals
  • Benzoquinones / metabolism
  • Biocatalysis
  • Biphenyl Compounds / metabolism
  • Cysteine / chemistry
  • Dihydroxyphenylalanine / analogs & derivatives
  • Dihydroxyphenylalanine / metabolism
  • Extracellular Matrix Proteins / chemistry
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / isolation & purification
  • Extracellular Matrix Proteins / metabolism*
  • Hydrogen-Ion Concentration
  • Hydrophobic and Hydrophilic Interactions
  • Models, Molecular*
  • Mytilus / physiology*
  • Nuclear Magnetic Resonance, Biomolecular
  • Oxidation-Reduction
  • Picrates / metabolism
  • Protein Conformation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Benzoquinones
  • Biphenyl Compounds
  • Extracellular Matrix Proteins
  • Picrates
  • Recombinant Proteins
  • Dihydroxyphenylalanine
  • dopaquinone
  • 1,1-diphenyl-2-picrylhydrazyl
  • Cysteine