Incorporation of Human Growth Hormone-2 into Proteoliposome Enhances Tissue Regeneration with Anti-Oxidant and Anti-Senescence Activities

Title
Incorporation of Human Growth Hormone-2 into Proteoliposome Enhances Tissue Regeneration with Anti-Oxidant and Anti-Senescence Activities
Author(s)
조경현이은영김소희
Keywords
HIGH-DENSITY-LIPOPROTEIN; APOLIPOPROTEIN-A-I; GESTATIONAL DIABETES-MELLITUS; HUMAN-PREGNANCY; PLACENTAL GH; FETAL-GROWTH; LIPID-FREE; APOA-I; PROTEINS; SECRETION
Issue Date
201502
Publisher
MARY ANN LIEBERT, INC
Citation
REJUVENATION RESEARCH, v.18, no.1, pp.20 - 29
Abstract
Human growth hormone-2 (GH-2) is a 191-amino-acid protein also known as human placental hormone. During pregnancy, continuous secretion of GH-2 appears to have important implications for physiological adjustment to gestation, especially in controlling levels of maternal insulin-like growth factor 1. To compare the physiological activity of GH-2 between lipid-free and lipid-bound states, GH-2 was expressed and incorporated into proteoliposome. GH-2 was expressed and purified using a pET28(a)-GH-2 vector in an Escherichia coli system. Purified GH-2 was then characterized and synthesized into reconstituted high-density lipoprotein (rHDL). The expression yield of GH-2 was 20-30 mg by BL21 (DE3) cells in 1 liter of Luria-Bertani broth. Purified GH-2 of at least 98% purity (23 kDa) was incorporated into rHDL with human apolipoprotein A-I (ApoA-I) and palmitoyloleoyl phosphatidylcholine (POPC) at a 1:1:95 (GH-2:ApoA-I:POPC) molar ratio. Structural analysis revealed that GH-2 had a 44% alpha-helix content and a wavelength maximum fluorescence (WMF) of 349 nm in a lipid-free state. In a lipid-bound state, the WMF of GH-2 was similar to 4 nm blue-shifted (345 nm), with 50% of alpha-helix content. The lipid-bound GH-2 showed enhanced anti-atherosclerotic activity and anti-senescence activity with inhibition of fructose-mediated glycation. A fin regeneration experiment using zebrafish (17 weeks old, n=9) showed that lipid-bound GH-2 enhanced regeneration efficiency by 44% compared to native GH-2 (in the lipid-free state) without any notable side effects. GH-2 has anti-oxidant activity to enhance tissue regeneration as well as to exert anti-diabetic activity. Incorporation of GH-2 into rHDL can enhance structural stability and tissue regeneration efficiency in vertebrate models, indicating a synergetic effect between GH-2 and ApoA-I in rHDL.
URI
http://hdl.handle.net/YU.REPOSITORY/33513http://dx.doi.org/10.1089/rej.2014.1594
ISSN
1549-1684
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