Human-derived fusogenic peptides for the intracellular delivery of proteins

Kei Sudo, Keisuke Niikura, Kouta Iwaki, Shunshi Kohyama, Kei Fujiwara, Nobuhide Doi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The cytosolic delivery of therapeutic proteins (e.g., antibodies or enzymes) by cell-penetrating peptides (CPPs), such as a human immunodeficiency virus-derived TAT peptide, is facilitated by fusogenic peptides (FPs). For instance, we recently demonstrated that an FP, B18, which is derived from a sea urchin gamete fusion protein, promotes endosomal escape of an enhanced green fluorescent protein (eGFP)-TAT fusion protein directly conjugated to it. However, the potential clinical use of FPs raises concerns because all conventional FPs are non-human-derived. To solve this problem, we have attempted to identify novel human-derived FPs from two human proteins, including a human sperm protein, IZUMO1, which is involved in gamete recognition and fusion, and a human endogenous retroviral envelope protein, Syncytin1, which is involved in placental morphogenesis. Partial peptides from the core domains of the abovementioned proteins were chosen as candidates to generate human-derived FPs. We prepared fusion proteins of these peptides with eGFP and TAT in Escherichia coli and observed the localization of these fusion proteins in HeLa cells using confocal microscopy. Our results suggested that a 19-residues peptide of Syncytin1 (positions 322–340), named S19, possessed strong intracellular uptake activities with no detectable cytotoxicity. In addition, we estimated the number of molecules that escaped from endosomes using a nuclear localization signal, suggesting that the S19 peptide stimulated the intracellular delivery of TAT-fused eGFP by ~90-fold. Furthermore, we confirmed that S19 promoted the intracellular delivery of eGFP to various human cell lines, including HeLa, A431, HepG2, and SK-N-SH. In addition, we demonstrated that not only eGFP but also SNAP-tag and β-galactosidase were delivered efficiently and retained their activities.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalJournal of Controlled Release
Volume255
DOIs
Publication statusPublished - 2017 Jun 10

Fingerprint

Peptides
Proteins
Germ Cells
Galactosidases
Cell-Penetrating Peptides
Nuclear Localization Signals
Sea Urchins
Endosomes
Morphogenesis
HeLa Cells
Confocal Microscopy
Spermatozoa
enhanced green fluorescent protein
HIV
Escherichia coli
Cell Line
Antibodies
Enzymes

Keywords

  • Cell-penetrating peptides
  • Endocytosis
  • IZUMO1
  • Syncytin1

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Human-derived fusogenic peptides for the intracellular delivery of proteins. / Sudo, Kei; Niikura, Keisuke; Iwaki, Kouta; Kohyama, Shunshi; Fujiwara, Kei; Doi, Nobuhide.

In: Journal of Controlled Release, Vol. 255, 10.06.2017, p. 1-11.

Research output: Contribution to journalArticle

Sudo, Kei ; Niikura, Keisuke ; Iwaki, Kouta ; Kohyama, Shunshi ; Fujiwara, Kei ; Doi, Nobuhide. / Human-derived fusogenic peptides for the intracellular delivery of proteins. In: Journal of Controlled Release. 2017 ; Vol. 255. pp. 1-11.
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