Mass spectrometric mapping of fibrinogen conformations at poly(ethylene terephthalate) interfaces.
We have characterized the adsorption of bovine fibrinogen onto the biomedical polymer polyethylene terephthalate (PET) by performing mass spectrometric mapping with a lysine-reactive biotin label. After digestion with trypsin, MALDI-TOF mass spectrometry was used to detect peptides from biotinylated bovine fibrinogen, with the goal of identifying lysines that were more accessible for reaction with the chemical label after adsorption. Peptides within domains that are believed to contribute to heparin binding, leukocyte activation, and platelet adhesion were found to be biotin labeled only after bovine fibrinogen adsorbed to the PET surface. Additionally, the accessibility of lysine residues throughout the entire molecule was observed to increase as the concentration of the adsorbing bovine fibrinogen solution decreased, suggesting that the proximity of biologically active motifs to hydrophilic residues leads to their exposure. The surface area per adsorbed bovine fibrinogen molecule was quantified on PET using optical waveguide lightmode spectroscopy (OWLS), which revealed higher surface densities for bovine fibrinogen adsorbed from higher concentration solutions. By measuring changes in both the identity and conformation of proteins that adsorb from complex mixtures such as blood or plasma, this technique may have applications in fundamental studies of protein adsorption and may allow for more accurate predictions of the biocompatibility of materials.
Scott
Evan A
EA
Department of Biomedical Engineering, Washington University, Campus Box 1097, One Brookings Drive, Saint Louis, MO 63130, USA.
Elbert
Donald L
DL
D000327:Adsorption; D001665:Binding Sites; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D005340:Fibrinogen / Q000737:chemistry* / Q000648:ultrastructure*; D013058:Mass Spectrometry / Q000379:methods*; D008422:Materials Testing / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*; D011093:Polyethylene Terephthalates; D011485:Protein Binding; D011487:Protein Conformation; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D011093:Polyethylene Terephthalates; D011092:Polyethylene Glycols; D005340:Fibrinogen