期刊
CHEMBIOCHEM
卷 13, 期 12, 页码 1818-1825出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201100792
关键词
cleavage; crosslinking; labeling; protein derivatization; quinones
资金
- NIH [GM054403]
- Howard Hughes Medical Institute international research fellowship
We show that mushroom tyrosinase catalyzes the formation of reactive o-quinones on unstructured, tyrosine-rich sequences such as hemagglutinin (HA) tags (YPYDVPDYA). In the absence of exogenous nucleophiles and at low protein concentrations, the o-quinone decomposes with fragmentation of the HA tag. At higher protein concentrations (>5 mg?mL-1), crosslinking is observed. Besthorn's reagent intercepts the o-quinone to give a characteristic pink complex that can be observed directly on a denaturing SDS-PAGE gel. Similar labeled species can be formed by using other nucleophiles such as Cy5-hydrazide. These reactions are selective for proteins bearing HA and other unstructured poly-tyrosine-containing tags and can be performed in lysates to create specifically tagged proteins.
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