An attempt to modulate the microporous diffusion of a model polypeptide by altering its secondary structure

Drug Deliv. 2003 Apr-Jun;10(2):65-72. doi: 10.1080/713840357.

Abstract

The purpose of the present study was to determine whether intentional alteration of the secondary structure of a model polypeptide, conantokin-G, influenced the rate and extent of aqueous pore diffusion across a synthetic microporous membrane. Use of a microporous synthetic membrane allowed for analysis of polypeptide transport without the confounding variables of protein binding, acid- and/or enzyme-mediated degradation, endocytotic uptake, and enzymatic inactivation associated with a biological membrane. Conantokin-G was intentionally changed from its native random coil structure to the alpha-helix structure using calcium, and both structures were verified using circular dichroism. The alpha-helix structure of conantokin-G was retained even after additional free calcium was removed by equilibrium dialysis. Over the concentration range of 1.25 to 20 mM, there was a linear relationship between the solution calcium concentration and the percent of the alpha-helix conformer present. The apparent permeability, the apparent aqueous diffusion coefficient with and without inclusion of the Renkin function, and the hydrodynamic radii estimated by diffusion and a computer-software program were calculated for the random coil and alpha-helix structures of conantokin-G. Calcium-mediated conversion of conantokin-G to its alpha-helix structure did not significantly (p >.05) change its apparent permeability across a microporous membrane. It is suggested that perhaps complete conversion to the alpha-helix structure of only a fraction of the conantokin-G molecules (only 0.45 or 45% of the molecules can be converted to the alpha-helix structure at Ca(2+) concentrations >or= 20 mM) may have limited the extent of transport of the alpha-helix conformer.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chemistry, Pharmaceutical / methods*
  • Conotoxins / chemistry*
  • Conotoxins / metabolism
  • Diffusion / drug effects*
  • Dosage Forms
  • Membranes, Artificial
  • Micropore Filters
  • Peptides* / chemical synthesis
  • Peptides* / chemistry*
  • Permeability / drug effects
  • Protein Structure, Secondary*
  • Snails
  • Structure-Activity Relationship

Substances

  • Conotoxins
  • Dosage Forms
  • Membranes, Artificial
  • Peptides
  • conotoxin GV