期刊
ACS SYNTHETIC BIOLOGY
卷 3, 期 6, 页码 353-362出版社
AMER CHEMICAL SOC
DOI: 10.1021/sb400116x
关键词
cell-free protein synthesis; non-natural amino acid; Escherichia coli; Click chemistry; protein polymerization; active biomaterials
资金
- Global Climate and Energy Project (GCEP) at Stanford Univercity
We report the synthesis of active polymers of superfolder green fluorescent protein (sfGFP) in one step decamer using Click chemistry. Up to six copies of the non-natural amino acids (nnAAs) p-azido-L-phenylalanine (pAzF) or pp-ropargyloxy-L-phenylalanine (pPaF) were site-specifically inserted into sfGFP by cell-free protein synthesis (CFPS). sfGFP containing two or three copies of these nnAAs were monomer coupled by copper-catalyzed azide alkyne cycloaddition to synthesize linear or branched protein polymers, respectively. The protein polymers retained >= 63% of their specific activity (i.e., fluorescence) after coupling. Polymerization of a concentrated solution of triply substituted sfGFP resulted in fluorescent macromolecular particles. Our method can be generalized to synthesize polymers of a protein or copolymers of any two or more proteins, and the conjugation sites can be determined exactly by standard genetic manipulation. Polymers of proteins and small molecules can also be created with this technology to make a new class of scaffolds or biomaterials.
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