title: Cell membrane penetrating function of the nuclear localization sequence in human cytokine IL-1α.
abstract: Cytokines are released from the cell, bind to their receptors, and affect cellular responses. The precursor form of interleukin 1 alpha (pIL-1α) has a nuclear localization sequence (NLS) that causes it to be localized to the nucleus and regulate specific gene expression. The amino acids of the NLS are basic amino acid-rich sequences, as is the cell penetrating peptide (CPP), which has been widely studied as a way to deliver macromolecules into cells. Here, we hypothesized that the NLS in pIL-1α (pIL-1αNLS) can penetrate the cell membrane and it could deliver macromolecules such as protein in vivo. We characterized cell membrane penetration ability of pIL-1αNLS or its tandem repeated form (2pIL-1αNLS) to enhance its intracellular delivery efficiency. 2pIL-1αNLS showed comparable protein delivery efficiency to TAT-CPP and it mediates endocytosis following heparan sulfate interaction. 2pIL-1αNLS conjugated enhanced green fluorescence protein was localized to the nucleus and the cytoplasm. Intra-peritoneal administration of 2pIL-1αNLS conjugated dTomato protein showed remarkable in vivo intracellular delivery efficiency in various tissues including spleen, liver, and intestine in mice. Moreover, cytotoxicity of 2pIL-1αNLS was not observed even at 100 μM. Our results demonstrate cell membrane-penetrating function of NLS in pIL-1α, which can be used as a safe therapeutic macromolecular delivery peptide.
authors.affiliation: Department of Life Science, Research Institute for Natural Sciences, Hanyang University, Seoul, 133-791, Republic of Korea.
authors.forename: Ja-Hyun
authors.initials: JH
authors.lastname: Koo
authors.forename: Heeseok
authors.initials: H
authors.lastname: Yoon
authors.forename: Won-Ju
authors.initials: WJ
authors.lastname: Kim
authors.forename: Sangho
authors.initials: S
authors.lastname: Lim
authors.forename: Hong-Jai
authors.initials: HJ
authors.lastname: Park
authors.forename: Je-Min
authors.initials: JM
authors.lastname: Choi
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D053582:Interleukin-1alpha / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019169:Jurkat Cells; D051379:Mice; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; C508584:IL1A protein, human; D053582:Interleukin-1alpha; D019913:Nuclear Localization Signals; D010446:Peptide Fragments; D011994:Recombinant Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus