A novel model system for design of biomaterials based on recombinant analogs of spider silk proteins

J Neuroimmune Pharmacol. 2009 Mar;4(1):17-27. doi: 10.1007/s11481-008-9129-z. Epub 2008 Oct 7.

Abstract

Spider dragline silk possesses impressive mechanical and biochemical properties. It is synthesized by a couple of major ampullate glands in spiders and comprises of two major structural proteins--spidroins 1 and 2. The relationship between structure and mechanical properties of spider silk is not well understood. Here, we modeled the complete process of the spider silk assembly using two new recombinant analogs of spidroins 1 and 2. The artificial genes sequence of the hydrophobic core regions of spidroin 1 and 2 have been designed using computer analysis of existing databases and mathematical modeling. Both proteins were expressed in Pichia pastoris and purified using a cation exchange chromatography. Despite the absence of hydrophilic N- and C-termini, both purified proteins spontaneously formed the nanofibrils and round micelles of about 1 microm in aqueous solutions. The electron microscopy study has revealed the helical structure of a nanofibril with a repeating motif of 40 nm. Using the electrospinning, the thin films with an antiparallel beta-sheet structure were produced. In summary, we were able to obtain artificial structures with characteristics that are perspective for further biomedical applications, such as producing three-dimensional matrices for tissue engineering and drug delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials / chemistry*
  • Circular Dichroism
  • Microscopy, Atomic Force
  • Microscopy, Electron, Scanning
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Models, Statistical
  • Nanotechnology
  • Recombinant Proteins / chemistry
  • Silk / chemistry*
  • Silk / genetics*
  • Silk / ultrastructure
  • Solutions
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrophotometry, Infrared
  • Spiders / chemistry*
  • Spiders / genetics*
  • Tissue Engineering

Substances

  • Biocompatible Materials
  • Recombinant Proteins
  • Silk
  • Solutions