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In Situ and Timed Extraction of Cellular Peptides from Live HeLa Cells by Photo-Switchable Mesoporous Silica Nanocarriers
journal contribution
posted on 2016-08-08, 00:00 authored by Yi Zhang, Jiaxi Peng, Zheyi Liu, Hanfa Zou, Ren’an WuIn situ and timed extraction of cellular
peptides is a great challenge
for dynamic and global proteomic investigation of live cells. In this
work, a mesoporous silica nanocarrier with photoswitchable off/on
coumarin gates (MSNcg) was developed for capturing peptides from the
cytosol of living HeLa cells. The MSNcg was constructed from mesoporous
silica nanoparticle (MSN) and its subsequent modifications with TAT
peptides and coumarin, to endow the features of the size-exclusion
effect of the mesoporous silica and the localization of nanocarrier
at cytosol by TAT peptide and to control the closing and opening of
the coumarin gates by reversible photodimerization and photocleavage.
With the pre-endocytosing of MSNcg, 126 cytosol peptides were harvested
and identified from living HeLa cells. Moreover, 3 peptides were captured
containing dynamic and changeable information. The extraction strategy
of using MSNcg exhibited promising potentials in the in situ and dynamic
extraction of endogenous peptides and/or proteins from living systems.