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Incorporation of human growth hormone-2 into proteoliposome enhances tissue regeneration with anti-oxidant and anti-senescence activities.

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% α-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 ∼4 nm blue-shifted (345 nm), with 50% of α-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.

journal_name

Rejuvenation Res

journal_title

Rejuvenation research

authors

Kim SH,Lee EY,Cho KH

doi

10.1089/rej.2014.1594

subject

Has Abstract

pub_date

2015-02-01 00:00:00

pages

20-9

issue

eissn

1549-1684

issn

1557-8577

journal_volume

pub_type

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Incorporation of human growth hormone-2 into proteoliposome enhances tissue regeneration with anti-oxidant and anti-senescence activities.