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Site-specific incorporation of phosphotyrosine using an expanded genetic code.

ABSTRACT: Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phosphorylation in negative regulation of ubiquitination.

SUBMITTER: Hoppmann C 

PROVIDER: S-EPMC5577362 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature

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Site-specific incorporation of phosphotyrosine using an expanded genetic code.

Hoppmann Christian C   Wong Allison A   Yang Bing B   Li Shuwei S   Hunter Tony T   Shokat Kevan M KM   Wang Lei L  

Nature chemical biology 20170612 8

Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phos  ...[more]

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