ac6b02853_si_001.pdf (1.66 MB)
High-Throughput Screening of Sulfated Proteins by Using a Genome-Wide Proteome Microarray and Protein Tyrosine Sulfation System
journal contribution
posted on 2017-02-17, 00:00 authored by Bo-Yu Huang, Po-Chung Chen, Bo-Han Chen, Chen-Chu Wang, Hsuan-Fu Liu, Yi-Zao Chen, Chien-Sheng Chen, Yuh-Shyong YangProtein
tyrosine sulfation (PTS) is a widespread posttranslational
modification that induces intercellular and extracellular responses
by regulating protein–protein interactions and enzymatic activity.
Although PTS affects numerous physiological and pathological processes,
only a small fraction of the total predicted sulfated proteins has
been identified to date. Here, we localized the potential sulfation
sites of Escherichia coli proteins
on a proteome microarray by using a 3′-phosphoadenosine 5′-phosphosulfate
(PAPS) synthase-coupled tyrosylprotein sulfotransferase (TPST) catalysis
system that involves in situ PAPS generation and TPST catalysis. Among
the 4256 E. coli K12 proteins, 875
sulfated proteins were identified using antisulfotyrosine primary
and Cy3-labeled antimouse secondary antibodies. Our findings add considerably
to the list of potential proteins subjected to tyrosine sulfation.
Similar procedures can be applied to identify sulfated proteins in
yeast and human proteome microarrays, and we expect such approaches
to contribute substantially to the understanding of important human
diseases.