Browsing by Author "Bloom, Adam"

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    Investigating the effect of proteoglycan 4 on hyaluronan solution properties using confocal fluorescence recovery after photobleaching
    (2014-08-26) Bloom, Adam; Schmidt, Tannin
    Hyaluronan (HA) and proteoglycan 4 (PRG4) are vital macromolecular constituents of synovial fluid (SF) that contribute synergistically to its biophysical properties. The objective of this thesis was to use confocal fluorescence recovery after photobleaching (FRAP) to determine whether PRG4 will decrease the permeability of simple HA solutions due to specific interactions between PRG4 and HA polymers in solution. Bovine and recombinant human PRG4 caused HA solutions of all concentrations to become less permeable to tracer molecules. This effect, specific to PRG4 and not seen with a control protein, was not dependent on protein structure as determined by reducing and alkylating PRG4. Additionally, confocal FRAP was successfully employed to study human SF samples of interest. These results demonstrate PRG4 can alter solution properties of HA, possibly due to entanglement through binding and/or HA crowding PRG4 into a self-assembled network. These results contribute to the understanding of PRG4’s role in SF.