Secondary structure analysis of proteins embedded in spherical polyelectrolyte brushes by FT-IR spectroscopy.
The adsorption of bovine serum albumin (BSA), bovine beta-lactoglobulin, and bovine pancreatic ribonuclease A onto spherical polyelectrolyte brushes (SPB) is reported. The SPB consist of narrowly distributed poly(styrene) core particles (diameter approximately 100 nm) onto which linear chains of anionic polyelectrolytes are grafted. The polyelectrolyte shell consists of either the weak polyelectrolyte poly(acrylic acid) or the strong polyacid poly(styrenesulfonate). The SPB particles are dispersed in H(2)O at room temperature. The secondary structure of the proteins was investigated by Fourier transform infrared spectroscopy in transmission mode before and during adsorption to these colloidal brushes. The alpha-helix and beta-sheet content of the proteins was nearly fully retained in the adsorbed state for all systems. Only in the case of BSA interacting with poly(styrenesulfonic) brushes could a slight loss of alpha-helix structure be observed. As the interaction of SPB and proteins can be controlled by the ionic strength in the buffer, additional experiments were performed to release the adsorbed protein. The amount of released protein was quantified and was found to be strongly dependent on the kind of protein and brush used. The secondary structure of the released proteins could be analyzed as well. An almost full preservation of secondary structure was found. This demonstrates that SPB are well-suited to immobilize proteins. The SPB can be charged and decharged under retention of the secondary structure of the biomolecules.
Wittemann
Alexander
A
Physikalische Chemie I, Universität Bayreuth, Universitaetsstrasse 30, D-95440 Bayreuth, Germany.
Ballauff
Matthias
M
D000818:Animals; D002417:Cattle; D004573:Electrolytes / Q000737:chemistry*; D007782:Lactoglobulins / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D004573:Electrolytes; D007782:Lactoglobulins; D011137:Polystyrenes; D011506:Proteins; D012710:Serum Albumin, Bovine; C003321:polystyrene sulfonic acid; D012259:Ribonuclease, Pancreatic