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ISSN
1672-9145
Acta Biochim Biophys Sin
2004, 36(9): 629–636
CN 31-1940/Q
A New Peptide with Membrane-permeable Function Derived from Human
Circadian Proteins
Tao PENG, Ying-Hui LIU, Chun-Lei YANG, Chao-Min WAN, Yao-Qi WANG,
and Zheng-Rong WANG*
Department of Biomedical Engineering, School of Basic Medicine and
Forensic, Sichuan University, Chengdu 610041, China
Abstract Basic peptides such as human immunodeficiency virus type 1 (HIV-1)
Tat-(48-60) and Drosophila Antennapedia-(43-58) have been reported to
have a membrane permeability and a carrier function for intracellular protein
delivery. Based on the fluorescence microscopic observations of the vascular
endothelial cells (ECV-304) and the primary cultured neuroglial cells, we found
that human Clock protein DNA-binding peptide [residue 35–47, hClock-(35-47)]
had a translocation activity very similar to Tat-(48-60). The cellular uptake
of hClock-(35-47) increases with the increase of incubation time and concentration.
The internalization effect at 4 °C was same as that at 37 °C. Internalization
of hClock-(35-47) was saturable and could be inhibited by the excess of the
other MPPs. Moreover, the uptake of these peptides were significantly inhibited
in the presence of heparan sulfate. These results strongly suggested that the
hClock-(35-47) shared a common or very similar internalization pathway with
other MPPs. Furthermore, we injected rat through the common carotid artery with
hClock-(35-47)-FITC peptide, and cryostat sections of the brain were prepared
and observed using a fluorescence microscope. Result showed that the peptide
had the ability to translocate through the blood-brain barrier. It is promising
to provide a new safe carrier for the intracellular and encephalic treatment.
Key words membrane-permeable peptide
(MPP); cell membrane; hClock; DNA-binding peptide; blood-brain barrier
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Received: April 1,
2004
Accepted: July 12, 2004
This work was supported
by the grants from the National Natural Science Foundation of China (No.
39970275, 30070278, No. 30070198)
*Corresponding author: Tel, 86-28-85502051; Fax, 86-28-85503204;
E-mail, [email protected]