Nontoxic membrane translocation peptide from protamine, low molecular weight protamine (LMWP), for enhanced intracellular protein delivery: in vitro and in vivo study.
Naturally derived, nontoxic peptides from protamine by the authors, termed low molecular weight protamines (LMWPs), possess high arginine content and carry significant sequence similarity to that of TAT, by far the most potent protein transduction domain peptide. Therefore, it was hypothesized that these LMWPs would also inherit the similar translocation activity across the cell membrane, which enables any impermeable species to be transduced into the cells. LMWPs were prepared by enzymatic digestion of protamine, examined their capability of transducing an impermeable protein toxin into the tumor cells by chemical conjugation, and determined cytotoxicity of transduced protein toxin (e.g., gelonin) against cancer cell lines and a tumor-bearing mouse. In vitro results showed that LMWPs could indeed translocate themselves into several mammalian cell lines as efficiently as TAT, thereby transducing impermeable gelonin into the cells by chemical conjugation. In vivo studies further confirmed that LMWP could carry an impermeable gelonin across the tumor mass and subsequently inhibit the tumor growth. In conclusion, the presence of equivalent cell translocation potency, absence of toxicity of peptide itself, and the suitability for low-cost production by simple enzymatic digestion could expand the range of clinical applications of LMWPs, including medical imaging and gene/protein therapies.
Park
Yoon Jeong
YJ
Craniomaxillofacial Reconstructive Science Major, School of Dentistry and Intellectual Biointerface Engineering Center, Seoul National University, Seoul, South Korea. parkyj@snu.ac.kr
Chang
Li-Chien
LC
Liang
Jun Feng
JF
Moon
Cheol
C
Chung
Chong-Pyoung
CP
Yang
Victor C
VC
D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D015696:Gene Products, tat / Q000494:pharmacology; D006801:Humans; D026901:Membrane Transport Proteins / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000188:drug therapy; D010446:Peptide Fragments / Q000494:pharmacology*; D010940:Plant Proteins / Q000494:pharmacology; D011479:Protamines / Q000494:pharmacology* / Q000627:therapeutic use; D021381:Protein Transport; D054789:Ribosome Inactivating Proteins, Type 1
D015696:Gene Products, tat; D026901:Membrane Transport Proteins; D010446:Peptide Fragments; D010940:Plant Proteins; D011479:Protamines; D054789:Ribosome Inactivating Proteins, Type 1; C025674:GEL protein, Gelonium multiflorum