Glycosaminoglycan-mediated coacervation of tropoelastin abolishes the critical concentration, accelerates coacervate formation, and facilitates spherule fusion: implications for tropoelastin microassembly.
Elastogenesis and elastin repair depend on the secretion of tropoelastin from the cell, yet cellular production is low in the many biological systems that have been studied. To address the apparent paradox of a paucity of tropoelastin for cell surface microassembly, we examined the effects of the glycosaminoglycans heparin, heparan sulfate, and chondroitin sulfate B, on tropoelastin aggregate formation through coacervation. We found a significant effect, particularly of heparin, on the minimum or critical concentration of tropoelastin, which was required for microassembly, lowering critical concentration to a point that it was no longer detectable. The assemblies resulted in protein droplet formation that was visually indistinguishable from the spherules that typify coacervation. The spherules readily coalesced in the presence of heparin and higher concentrations of tropoelastin, resulting in an almost continuous layer of coacervated tropoelastin. Four stages of droplet behavior were observed: early droplet formation, approximately 6 mum droplet formation, and fusion of droplets followed by the formation of a coalesced layer. We conclude that glycosaminoglycans in the extracellular matrix have the capacity to promote coacervation at low concentrations of tropoelastin.
School of Molecular and Microbial Biosciences G08, University of Sydney, NSW 2006, Australia.
Yidong
Y
Tu
Anthony S
AS
Weiss
D003871:Dermatan Sulfate; D004549:Elastin; D005109:Extracellular Matrix / Q000737:chemistry; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin; D006497:Heparan Sulfate; D008954:Models, Biological; D011485:Protein Binding; D012996:Solutions; D000267:Tissue Adhesions; D014333:Tropoelastin / Q000737:chemistry*
D003871:Dermatan Sulfate; D004549:Elastin; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D012996:Solutions; D014333:Tropoelastin