Unexpected Multivalent Display of Proteins by Temperature Triggered Self-Assembly of Elastin-like Polypeptide Block Copolymers

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• 作者：Wafa Hassouneh ; Karl Fischer ; Sarah R. MacEwan ; Robert Branscheid ; Chuan Lawrence Fu ; Rihe Liu ; Manfred Schmidt ; Ashutosh Chilkoti
• 刊名：Biomacromolecules
• 出版年：2012
• 出版时间：May 14, 2012
• 年：2012
• 卷：13
• 期：5
• 页码：1598-1605
• 全文大小：375K
• 年卷期：v.13,no.5(May 14, 2012)
• ISSN：1526-4602

文摘

We report herein the unexpected temperature triggered self-assembly of proteins fused to thermally responsive elastin-like polypeptides (ELPs) into spherical micelles. A set of six ELP block copolymers (ELP_BC) differing in hydrophilic and hydrophobic block lengths were genetically fused to two single domain proteins, thioredoxin (Trx) and a fibronectin type III domain (Fn3) that binds the 伪_v尾₃ integrin. The self-assembly of these protein鈥揈LP_BC fusions as a function of temperature was investigated by UV spectroscopy, light scattering, and cryo-TEM. Self-assembly of the ELP_BC was unexpectedly retained upon fusion to the two proteins, resulting in the formation of spherical micelles with a hydrodynamic radius that ranged from 24 to 37 nm, depending on the protein and ELP_BC. Cryo-TEM images confirmed the formation of spherical particles with a size that was consistent with that measured by light scattering. The bioactivity of Fn3 was retained when presented by the ELP_BC micelles, as indicated by the enhanced uptake of the Fn3-decorated ELP_BC micelles in comparison to the unimer by cells that overexpress the 伪_v尾₃ integrin. The fusion of single domain proteins to ELP_BCs may provide a ubiquitous platform for the multivalent presentation of proteins.