Insights into the role of protein molecule size and structure on interfacial properties using designed sequences

Mirjana Dimitrijev-Dwyer, Lizhong He, Michael James, Andrew Nelson, Anton P J Middelberg

Research output: Contribution to journalArticleResearchpeer-review

12 Citations (Scopus)

Abstract

Mixtures of a large, structured protein with a smaller, unstructured component are inherently complex and hard to characterize at interfaces, leading to difficulties in understanding their interfacial behaviours and, therefore, formulation optimization. Here, we investigated interfacial properties of such a mixed system. Simplicity was achieved using designed sequences in which chemical differences had been eliminated to isolate the effect of molecular size and structure, namely a short unstructured peptide (DAMP1) and its longer structured protein concatamer (DAMP4). Interfacial tension measurements suggested that the size and bulk structuring of the larger molecule led to much slower adsorption kinetics. Neutron reflectometry at equilibrium revealed that both molecules adsorbed as a monolayer to the air-water interface (indicating unfolding of DAMP4 to give a chain of four connected DAMP1 molecules), with a concentration ratio equal to that in the bulk. This suggests the overall free energy of adsorption is equal despite differences in size and bulk structure. At small interfacial extensional strains, only molecule packing influenced the stress response. At larger strains, the effect of size became apparent, with DAMP4 registering a higher stress response and interfacial elasticity. When both componentswere present at the interface, most stress-dissipating movement was achieved by DAMP1. This work thus provides insights into the role of proteins' molecular size and structure on their interfacial properties, and the designed sequences introduced here can serve as effective tools for interfacial studies of proteins and polymers.

Original languageEnglish
Article number20120987
Number of pages12
JournalJournal of the Royal Society Interface
Volume10
Issue number80
DOIs
Publication statusPublished - 2013

Keywords

  • Adsorption
  • Interface
  • Neutron reflectometry
  • Peptide
  • Protein
  • Rheology

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