TY - JOUR
T1 - Carborane as an Alternative Efficient Hydrophobic Tag for Protein Degradation
AU - Asawa, Yasunobu
AU - Nishida, Kei
AU - Kawai, Kazuki
AU - Domae, Kiyotaka
AU - Ban, Hyun Seung
AU - Kitazaki, Akihiro
AU - Asami, Hiroya
AU - Kohno, Jun Ya
AU - Okada, Satoshi
AU - Tokuma, Hiraku
AU - Sakano, Daisuke
AU - Kume, Shoen
AU - Tanaka, Masaru
AU - Nakamura, Hiroyuki
N1 - Funding Information:
This work was partially supported by a Grant-in-Aid for Chemistry for Multimolecular Crowding Biosystems (20H04699 to H.N.) from MEXT, Japan, and JSPS KAKENHI (JP19H05720 to M.T. and 21J13988 to Y.A.) and performed under the Research Program of “Five-star Alliances” in “NJRC Mater. & Dev.”. The authors thank Prof. Minoru Ishikawa and Dr. Shusuke Tomoshige (Graduate School of Life Science, Tohoku University) for their valuable suggestions. H.S.B. thanks Five-star Alliance overseas researcher invitation program in “NJRC Mater. & Dev.”. Dedicated to the memory of late Professor M. Frederick Hawthorne (1928-2021).
Funding Information:
This work was partially supported by a Grant-in-Aid for Chemistry for Multimolecular Crowding Biosystems (20H04699 to H.N.) from MEXT Japan, and JSPS KAKENHI (JP19H05720 to M.T. and 21J13988 to Y.A.) and performed under the Research Program of ???Five-star Alliances in NJRC Mater. & Dev.
Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/11/17
Y1 - 2021/11/17
N2 - Carboranes 1 and 2 were designed and synthesized for hydrophobic tag (HyT)-induced degradation of HaloTag fusion proteins. The levels of the hemagglutinin (HA)-HaloTag2-green fluorescent protein (EGFP) stably expressed in Flp-In 293 cells were significantly reduced by HyT13, HyT55, and carboranes 1 and 2, with expression levels of 49, 79, 43, and 65%, respectively, indicating that carborane is an alternative novel hydrophobic tag (HyT) for protein degradation under an intracellular environment. To clarify the mechanism of HyT-induced proteolysis, bovine serum albumin (BSA) was chosen as an extracellular protein and modified with maleimide-conjugated m-carborane (MIC). The measurement of the ζ-potentials and the lysine residue modification with fluorescein isothiocyanate (FITC) of BSA-MIC conjugates suggested that the conjugation of carborane induced the exposure of lysine residues on BSA, resulting in the degradation via ubiquitin E3 ligase-related proteasome pathways in the cell.
AB - Carboranes 1 and 2 were designed and synthesized for hydrophobic tag (HyT)-induced degradation of HaloTag fusion proteins. The levels of the hemagglutinin (HA)-HaloTag2-green fluorescent protein (EGFP) stably expressed in Flp-In 293 cells were significantly reduced by HyT13, HyT55, and carboranes 1 and 2, with expression levels of 49, 79, 43, and 65%, respectively, indicating that carborane is an alternative novel hydrophobic tag (HyT) for protein degradation under an intracellular environment. To clarify the mechanism of HyT-induced proteolysis, bovine serum albumin (BSA) was chosen as an extracellular protein and modified with maleimide-conjugated m-carborane (MIC). The measurement of the ζ-potentials and the lysine residue modification with fluorescein isothiocyanate (FITC) of BSA-MIC conjugates suggested that the conjugation of carborane induced the exposure of lysine residues on BSA, resulting in the degradation via ubiquitin E3 ligase-related proteasome pathways in the cell.
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U2 - 10.1021/acs.bioconjchem.1c00431
DO - 10.1021/acs.bioconjchem.1c00431
M3 - Article
C2 - 34699716
AN - SCOPUS:85118716871
SN - 1043-1802
VL - 32
SP - 2377
EP - 2385
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 11
ER -