Coacervation characteristics of recombinant human tropoelastin.
Coacervation of soluble tropoelastin molecules is characterized by thermodynamically reversible association as temperature is increased under appropriately juxtaposed ionic conditions, protein concentration and pH. Coacervation plays a critical role in the assembly of these elastin precursors in elastic fiber formation. To examine the effect of physiological parameters on the ability of tropoelastin molecules to associate, solutions of recombinant human tropoelastin were monitored spectrophotometrically by light scattering over a broad range of temperatures. Coacervation of recombinant human tropoelastin is strongly influenced by the concentration of protein and NaCl and to a lesser extent on pH. Trends towards maximal association are apparent when each of these parameters is varied. Remarkably, optimal coacervation is found at 37 degrees C, 150 mM NaCl and pH 7-8. Using the data generated by time courses, estimates of thermodynamic parameters were made. These estimates confirm that coacervation is endothermic and is marked by a strong entropic contribution. Circular dichroism of recombinant human tropoelastin revealed that, rather than being random, the structure is compatible with being largely that, of an all-beta protein (with secondary structure estimated to be 3% alpha-helix, 41% beta-sheet, 21% beta-turn and 33% other), exhibiting a spectrum as previously seen for tropoelastin populations and soluble elastin from naturally-derived sources.
Vrhovski
B
B
Department of Biochemistry, University of Sydney, NSW, Australia.
Jensen
S
S
Weiss
A S
AS
D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009391:Nephelometry and Turbidimetry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry; D012965:Sodium Chloride / Q000494:pharmacology; D012995:Solubility; D013696:Temperature; D013816:Thermodynamics; D014333:Tropoelastin / Q000737:chemistry*
D011994:Recombinant Proteins; D014333:Tropoelastin; D012965:Sodium Chloride