Synthesis of recombinant high density lipoprotein with apolipoprotein A-I and apolipoprotein A-V

2011 ◽  
Vol 392 (5) ◽  
Author(s):  
Xinbo Zhang ◽  
Baosheng Chen

Abstract It has been shown that apolipoprotein A-V (apoA-V) over-expression significantly lowers plasma triglyceride levels and decreases atherosclerotic lesion development. To assess the feasibility of recombinant high density lipoprotein (rHDL) reconstituted with apoA-V and apolipoprotein A-I (apoA-I) as a therapeutic agent for hyperlipidemic disorder and atherosclerosis, a series of rHDL were synthesized in vitro with various mass ratios of recombinant apoA-I and apoA-V. It is interesting to find that apoA-V of rHDL had no effect on lipoprotein lipase (LPL) activation in vitro and very low density lipoprotein (VLDL) clearance in HepG2 cells and in vivo. By contrast, LPL activation and VLDL clearance were inhibited by the addition of apoA-V to rHDL. Furthermore, the apoA-V of rHDL could not redistribute from rHDL to VLDL after incubation at 37°C for 30 min. These findings suggest that an increase of apoA-V in rHDL could not play a role in VLDL clearance in vitro and in vivo, which could, at least in part, attribute to the lost redistribution of apoA-V from rHDL to VLDL and LPL binding ability of apoA-V in rHDL. The therapeutic application of rHDL reconstituted with apoA-V and apoA-I might need the construction of rHDL from which apoA-V could freely redistribute to VLDL.

2011 ◽  
Vol 31 (6) ◽  
pp. 1333-1341 ◽  
Author(s):  
Andrew J. Murphy ◽  
Kevin J. Woollard ◽  
Andreas Suhartoyo ◽  
Roslynn A. Stirzaker ◽  
James Shaw ◽  
...  

1999 ◽  
Vol 337 (3) ◽  
pp. 445-451 ◽  
Author(s):  
Moira A. CLAY ◽  
Daniel A. CEHIC ◽  
Diana H. PYLE ◽  
Kerry-Anne RYE ◽  
Philip J. BARTER

We have shown previously that apolipoprotein A (apoA)-I-containing high-density lipoprotein (HDL) particles are formed by the conjugation of lipid-free apoA-I with lipids derived from other lipoprotein fractions in a process dependent on non-esterified fatty acids, generated by the lipolysis of very-low-density lipoprotein (VLDL) or provided exogenously. In the present study, we show that this process is also able to generate HDL particles containing apoA-II (A-II HDL) and both apoA-I and apoA-II (A-I/A-II HDL). When lipid-free apoA-II was incubated with either VLDLs and lipoprotein lipase or LDLs and sodium oleate, a significant proportion of the apoA-II was recovered in the HDL density fraction. This was associated with the formation of several populations of HDL-sized particles with pre-β2 electrophoretic mobility, which contained phospholipids and unesterified cholesterol as their main lipid constituents. When both lipid-free apoA-I and lipid-free apoA-II were incubated with LDL and sodium oleate, both apolipoproteins were recovered in HDLs that contained phospholipids and unesterified cholesterol as their main lipids. Two populations of particles had diameters of 7.4 and 10.8 nm and pre-β2-migration; there was also a population of pre-β1-migrating particles of diameter 4.7 nm. ApoA-I and apoA-II were both present in the larger HDLs, whereas only apoA-I was present in the smaller particles. Immunoaffinity chromatography on an anti-(apoA-I)–Sepharose column revealed that 10–20% of the apoA-II resided in particles that also contained apoA-I. The majority of the A-I/A-II HDL were present in a population of pre-β2 particles of 10.8 nm diameter. These results in vitro illustrate a potential mechanism for the formation of HDLs containing both apoA-I and apoA-II.


Steroids ◽  
1990 ◽  
Vol 55 (7) ◽  
pp. 308-313
Author(s):  
I.J. Goldberg ◽  
R.S. Rosenfeld ◽  
I. Paul ◽  
L.K. Miller ◽  
M.L. Tiell

2017 ◽  
Vol 37 (1) ◽  
pp. 1
Author(s):  
Agus Slamet ◽  
Bayu Kanetro

Protein content of winged bean is almost the same as soybean, but the beany flavor is more poweful than soybean. Therefore the protein of winged bean was isolated prior to use as raw material of yogurt. This research was aimed to determine the potency of  hypocholestrolemic activity of yogurt protein isolate of winged  bean through in vivo bioassay by using Sprague Dawley male rats. The treatments of the research were yogurt feed treatment with concentration of yogurt 0 (standard feed without yogurt as a control), 2, and 4 g yogurt/day as low and high concentration treatment respectively for 4th weeks after hypercholesterol feed  treatment for 1 week. The blood lipid profile of rats, including triglyceride, cholesterol total, High Density Lipoprotein (HDL), Low  Density Lipoprotein (LDL) cholesterol were analysed on the 2nd  and 4th weeks for the yogurt feed treatment while for before  yogurt feed treatment, the evaluation were based on the  adaptation phase and the 1st week for hypercholesterol phase.  The result of this research showed that the blood triglyceride,  cholesterol total, LDL increased, and the blood HDL decreased in hypercholesterol phase before yogurt feed treatment. The potency of hypocholestrolemic of yogurt from protein isolate of winged  bean was shown by the decreasing of blood triglyceride,  cholesterol total, LDL and increasing the HDL cholesterol after the yogurt feed treatment with low and high concentration. That  indicated that yogurt that was made of protein isolate of winged  bean could reduced cholesterol. ABSTRAKBiji kecipir memiliki kadar protein yang hampir sama dengan  kedelai, namun bau langunya lebih tajam daripada kedelai,  sehingga perlu diisolasi proteinnya sebelum digunakan sebagai  bahan baku yogurt. Tujuan penelitian ini adalah menentukan  potensi hipokolesterolemik yogurt isolat proteun biji kecipir  melalui uji biologis in vivo menggunakan tikus jantan Sprague Dawley. Perlakuan penelitian ini adalah perlakuan pakan yogurt  dengan konsentrasi 0 (pakan standar tanpa penambahan yogurt sebagai kontrol), 2, dan 4 g yogurt/hari berturut-turut sebagai  perlakuan konsentrasi rendah dan tinggi selama 4 minggu  perlakuan pakan yogurt sesudah pemberian pakan hiperkolesterol selama 1 minggu. Profil lipida darah tikus meliputi kadar trigliserida, total kolesterol, kolesterol High Density  Lipoprotein (HDL), dan Low Density Lipoprotein (LDL) dianalisis  pada minggu ke 2 dan 4 minggu selama perlakuan pakan yogurt  dan sebelum perlakuan pakan yogurt yaitu pada fase pemeliharaan adaptasi dan 1 minggu pada fase pemeliharan  hiperkolesterol. Hasil penelitian ini menunjukkan bahwa  trigliserida, total kolesterol, dan kolesterol LDL meningkat dan kolesterol HDL menurun selama fase pemberian pakan  hiperkolesterol sebelum perlakuan pakan yogurt. Potensi  hipokolesterol yogurt isolat protein biji kecipir ditunjukkan dengan penurunan trigliserida, total kolesterol, dan kolesterol LDL, serta peningkatan kolesterol HDL sesudah perlakuan pakan yogurt dengan konsentrasi rendah maupun tinggi. Hal tersebut mengindikasikan bahwa yogurt isolat protein biji kecipir mampu menurunkan kolesterol.


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