TY - JOUR
T1 - Extending the Half-Life of a Protein in Vivo by Enzymatic Labeling with Amphiphilic Lipopeptides
AU - Takahara, Mari
AU - Mochizuki, Shinichi
AU - Wakabayashi, Rie
AU - Minamihata, Kosuke
AU - Goto, Masahiro
AU - Sakurai, Kazuo
AU - Kamiya, Noriho
N1 - Funding Information:
This work was financially supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI (No. 17H07320 to M.T., No. JP16H04581 to N.K., No. 18K12082 to S.M.). Part of this work was conducted at Kyushu University and supported by the Nanotechnology Platform Program (Molecule and Material Synthesis) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. The Asahi Glass Foundation also funded this project. We thank Prof. Takeshi Mori (Kyushu University) for advice on the retention of lipid/BSA complexes in vivo and the Edanz Group ( https://en-author-services.edanzgroup.com/ac ) for editing a draft of this manuscript.
Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/4/21
Y1 - 2021/4/21
N2 - Synthesis of lipid-protein conjugates is one of the significant techniques in drug delivery systems of proteins; however, the intact conjugation of a lipid and protein is yet challenging due to the hydrophobicity of lipid molecules. In order to facilitate easy handling of the lipid moiety in conjugation, we have focused on a microbial transglutaminase (MTG) that can ligate specific lysine (K) and glutamine (Q) residues in lipopeptides and a protein of interest. As MTG substrates, monolipid- and dilipid-fused amphiphilic short lipopeptide substrates (lipid-G3S-RHK or lipid2-KG3S-RHK) were designed. These amphiphilic lipopeptides and a model protein (enhanced green fluorescent protein, EGFP) fused with LLQG (LQ-EGFP) were both water-soluble, and thus lipid-protein conjugates were efficiently obtained through the MTG reaction with a >80% conversion rate of LQ-EGFP even using cholesterol-G3S-RHK. In vitro cell adhesion and in vivo half-life stability of the successfully obtained lipid-protein conjugates were evaluated, showing that the monocholesterol-G3S-RHK modification of a protein gave the highest cell adhesion efficiency and longest half-life time by formation of a stable albumin/lipid-protein complex.
AB - Synthesis of lipid-protein conjugates is one of the significant techniques in drug delivery systems of proteins; however, the intact conjugation of a lipid and protein is yet challenging due to the hydrophobicity of lipid molecules. In order to facilitate easy handling of the lipid moiety in conjugation, we have focused on a microbial transglutaminase (MTG) that can ligate specific lysine (K) and glutamine (Q) residues in lipopeptides and a protein of interest. As MTG substrates, monolipid- and dilipid-fused amphiphilic short lipopeptide substrates (lipid-G3S-RHK or lipid2-KG3S-RHK) were designed. These amphiphilic lipopeptides and a model protein (enhanced green fluorescent protein, EGFP) fused with LLQG (LQ-EGFP) were both water-soluble, and thus lipid-protein conjugates were efficiently obtained through the MTG reaction with a >80% conversion rate of LQ-EGFP even using cholesterol-G3S-RHK. In vitro cell adhesion and in vivo half-life stability of the successfully obtained lipid-protein conjugates were evaluated, showing that the monocholesterol-G3S-RHK modification of a protein gave the highest cell adhesion efficiency and longest half-life time by formation of a stable albumin/lipid-protein complex.
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U2 - 10.1021/acs.bioconjchem.1c00027
DO - 10.1021/acs.bioconjchem.1c00027
M3 - Article
C2 - 33689283
AN - SCOPUS:85103376452
SN - 1043-1802
VL - 32
SP - 655
EP - 660
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 4
ER -