Positively charged peptides can interact with each other, as revealed by solid phase binding assays

Anal Biochem. 2006 May 15;352(2):157-68. doi: 10.1016/j.ab.2006.03.002. Epub 2006 Mar 20.

Abstract

Solid phase assay systems such as enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), and overlay gels are used to study processes of protein-protein interactions. The common principle of all these methods is that they monitor the binding between soluble and surface-immobilized molecules. Following the use of bovine serum albumin (BSA)-peptide conjugates or isolated synthetic peptides and the above-mentioned solid phase assay systems, the results of the current work demonstrate that positively charged peptides can interact with each other. Both the ELISA and SPR methods demonstrated that the binding process reached saturation with K(d) values ranging between 1 and 14 nM. No interaction was observed between BSA conjugates bearing positively charged peptides and conjugates bearing negatively charged peptides or with pure BSA molecules, strengthening the view that interaction occurs only between positively charged peptides. However, interactions between peptides in solution were not observed by nuclear magnetic resonance (NMR) or by native gel electrophoresis. It appears that for positively charged molecules to interact, one of the binding partners must be immobilized to a surface, a process that may lead to the exposure of otherwise masked groups or atoms. We discuss the relevance of our findings for the use of solid phase assay systems to study interactions between biomolecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cattle
  • Enzyme-Linked Immunosorbent Assay / methods
  • Gels / chemistry
  • Nuclear Magnetic Resonance, Biomolecular
  • Peptides / chemistry*
  • Sensitivity and Specificity
  • Serum Albumin, Bovine / chemistry*
  • Static Electricity
  • Surface Plasmon Resonance / methods*
  • Surface Properties

Substances

  • Gels
  • Peptides
  • Serum Albumin, Bovine