Cross-linking by protein oxidation in the rapidly setting gel-based glues of slugs

J Exp Biol. 2011 May 15;214(Pt 10):1699-706. doi: 10.1242/jeb.051581.

Abstract

The terrestrial slug Arion subfuscus secretes a glue that is a dilute gel with remarkable adhesive and cohesive strength. The function of this glue depends on metals, raising the possibility that metal-catalyzed oxidation plays a role. The extent and time course of protein oxidation was measured by immunoblotting to detect the resulting carbonyl groups. Several proteins, particularly one with a relative molecular mass (M(r)) of 165 x 10³, were heavily oxidized. Of the proteins known to distinguish the glue from non-adhesive mucus, only specific size variants were oxidized. The oxidation appears to occur within the first few seconds of secretion. Although carbonyls were detected by 2,4-dinitrophenylhydrazine (DNPH) in denatured proteins, they were not easily detected in the native state. The presence of reversible cross-links derived from carbonyls was tested for by treatment with sodium borohydride, which would reduce uncross-linked carbonyls to alcohols, but stabilize imine bonds formed by carbonyls and thus lead to less soluble complexes. Consistent with imine bond formation, sodium borohydride led to a 20-35% decrease in the amount of soluble protein with a M(r) of 40-165 (x 10³) without changing the carbonyl content per protein. In contrast, the nucleophile hydroxylamine, which would competitively disrupt imine bonds, increased protein solubility in the glue. Finally, the primary amine groups on a protein with a M(r) of 15 x 10³ were not accessible to acid anhydrides. The results suggest that cross-links between aldehydes and primary amines contribute to the cohesive strength of the glue.

Publication types

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

MeSH terms

  • Adhesives / chemistry
  • Adhesives / metabolism*
  • Aldehydes / chemistry
  • Aldehydes / metabolism
  • Amines / chemistry
  • Amines / metabolism
  • Animals
  • Borohydrides
  • Cross-Linking Reagents / chemistry
  • Cross-Linking Reagents / metabolism*
  • Gastropoda / chemistry*
  • Gels / chemistry
  • Gels / metabolism*
  • Hydrazines
  • Hydroxylamine
  • Immunoblotting
  • Oxidation-Reduction
  • Proteins / metabolism*

Substances

  • Adhesives
  • Aldehydes
  • Amines
  • Borohydrides
  • Cross-Linking Reagents
  • Gels
  • Hydrazines
  • Proteins
  • dinitrophenylhydrazine
  • Hydroxylamine
  • sodium borohydride