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Dimerization of 30Kc19 protein in the presence of amphiphilic moiety and importance of Cys‐57 during cell penetration
Author(s) -
Park Hee Ho,
Sohn Youngsoo,
Yeo Ji Woo,
Park Ju Hyun,
Lee Hong Jai,
Ryu Jina,
Rhee Won Jong,
Park Tai Hyun
Publication year - 2014
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201400253
Subject(s) - amphiphile , phospholipid , lipid bilayer , biochemistry , chemistry , cell penetrating peptide , penetration (warfare) , moiety , cell , biophysics , liposome , cell membrane , membrane , biology , stereochemistry , organic chemistry , operations research , engineering , copolymer , polymer
Abstract Recently, the recombinant 30Kc19 protein, originating from silkworm hemolymph of Bombyx mori has attracted attention due to its cell‐penetrating property and potential application as a protein delivery system. However, this observation of penetration across cell membrane has raised questions concerning the interaction of the protein‐lipid bilayer. Here, we report a dimerization propensity of the 30Kc19 protein in the presence of amphiphilic moieties; sodium dodecyl sulfate (SDS) or phospholipid. Native PAGE showed that the 30Kc19 monomer formed a dimer when SDS or phospholipid was present. In the glutathione‐ S ‐transferase (GST) pull‐down assay, supplementation of the 30Kc19 protein to mammalian cell culture medium showed dimerization and penetration; due to phospholipids at the cell membrane, the main components of the lipid bilayer. Mutagenesis was performed, and penetration was observed by 30Kc19 C76A and not 30Kc19 C57A, which meant that the presence of cysteine at position 57 (Cys‐57) is involved in dimerization of the 30Kc19 at the cell membrane during penetration. We anticipate application of the native 30Kc19 protein with high cell‐penetrating efficiency for delivery of cargos to various cell types. The intracellular cargo delivery using the 30Kc19 protein is a non‐virus derived (e.g. TAT) delivery method, which can open up new approaches for the delivery of therapeutics in bioindustries, such as pharma‐ and cosmeceuticals.