scholarly journals Structural and Functional Changes of Reconstituted High-Density Lipoprotein (HDL) by Incorporation of α-synuclein: A Potent Antioxidant and Anti-Glycation Activity of α-synuclein and apoA-I in HDL at High Molar Ratio of α-synuclein

Molecules ◽  
2021 ◽  
Vol 26 (24) ◽  
pp. 7485
Author(s):  
Kyung-Hyun Cho

α-synuclein (α-syn) is a major culprit of Parkinson’s disease (PD), although lipoprotein metabolism is very important in the pathogenesis of PD. α-syn was expressed and purified using the pET30a expression vector from an E. coli expression system to elucidate the physiological effects of α-syn on lipoprotein metabolism. The human α-syn protein (140 amino acids) with His-tag (8 amino acids) was expressed and purified to at least 95% purity. Isoelectric focusing gel electrophoresis showed that the isoelectric point (pI) of α-syn and apoA-I were pI = 4.5 and pI = 6.4, respectively. The lipid-free α-syn showed almost no phospholipid-binding ability, while apoA-I showed rapid binding ability with a half-time (T1/2) = 8 ± 0.7 min. The α-syn and apoA-I could be incorporated into the reconstituted HDL (rHDL, molar ratio 95:5:1:1, palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC):cholesterol:apoA-I:α-syn with the production of larger particles (92 Å) than apoA-I-rHDL (86 and 78 Å) and α-syn-rHDL (65 Å). An rHDL containing both apoA-I and α-syn showed lower α-helicity around 45% with a red shift of the Trp wavelength maximum fluorescence (WMF) from 339 nm, while apoA-I-HDL showed 76% α-helicity and 337 nm of WMF. The denaturation by urea addition showed that the incorporation of α-syn in rHDL caused a larger increase in the WMF than apoA-I-rHDL, suggesting that the destabilization of the secondary structure of apoA-I by the addition of α-syn. On the other hand, the addition of α-syn induced two-times higher resistance to rHDL glycation at apoA-I:α-syn molar ratios of 1:1 and 1:2. Interestingly, low α-syn in rHDL concentrations, molar ratio of 1:0.5 (apoA-I:α-syn), did not prevent glycation with more multimerization of apoA-I. In the lipid-free and lipid-bound state, α-syn showed more potent antioxidant activity than apoA-I against cupric ion-mediated LDL oxidation. On the other hand, microinjection of α-syn (final 2 μM) resulted in 10% less survival of zebrafish embryos than apoA-I. A subcutaneous injection of α-syn (final 34 μM) resulted in less tail fin regeneration than apoA-I. Interestingly, incorporation of α-syn at a low molar ratio (apoA-I:α-syn, 1:0.5) in rHDL resulted destabilization of the secondary structure and impairment of apoA-I functionality via more oxidation and glycation. However, at a higher molar ratio of α-syn in rHDL (apoA-I:α-syn = 1:1 or 1:2) exhibited potent antioxidant and anti-glycation activity without aggregation. In conclusion, there might be a critical concentration of α-syn and apoA-I in HDL-like complex to prevent the aggregation of apoA-I via structural and functional enhancement.

2010 ◽  
Vol 10 (1) ◽  
pp. 1-6 ◽  
Author(s):  
R. Murillo ◽  
J. Sarasa ◽  
M. Lanao ◽  
J. L. Ovelleiro

The degradation of chlorpyriphos by different advanced oxidation processes such as photo-Fenton, TiO2, TiO2/H2O2, O3 and O3/H2O2 was investigated. The photo-Fenton and TiO2 processes were optimized using a solar chamber as light source. The optimum dosages of the photo-Fenton treatment were: [H2O2]=0.01 M; [Fe3 + ]=10 mg l−1; initial pH = 3.5. With these optimum conditions total degradation was observed after 15 minutes of reaction time. The application of sunlight was also efficient as total degradation was achieved after 60 minutes. The optimum dosage using only TiO2 as catalyst was 1,000 mg l−1, obtaining the maximum degradation at 20 minutes of reaction time. On the other hand, the addition of 0.02 M of H2O2 to a lower dosage of TiO2 (10 mg l−1) provides the same degradation. The ozonation treatment achieved complete degradation at 30 minutes of reaction time. On the other hand, it was observed that the degradation was faster by adding H2O2 (H2O2/O3 molar ratio = 0.5). In this case, total degradation was observed after 20 minutes.


1948 ◽  
Vol 21 (4) ◽  
pp. 853-859
Author(s):  
R. F. A. Altman

Abstract As numerous investigators have shown, some of the nonrubber components of Hevea latex have a decided accelerating action on the process of vulcanization. A survey of the literature on this subject points to the validity of certain general facts. 1. Among the nonrubber components of latex which have been investigated, certain nitrogenous bases appear to be most important for accelerating the rate of vulcanization. 2. These nitrogen bases apparently occur partly naturally in fresh latex, and partly as the result of putrefaction, heating, and other decomposition processes. 3. The nitrogen bases naturally present in fresh latex at later stages have been identified by Altman to be trigonelline, stachhydrine, betonicine, choline, methylamine, trimethylamine, and ammonia. These bases are markedly active in vulcanization, as will be seen in the section on experimental results. 4. The nitrogenous substances formed by the decomposition processes have only partly been identified, on the one hand as tetra- and pentamethylene diamine and some amino acids, on the other hand as alkaloids, proline, diamino acids, etc. 5. It has been generally accepted that these nitrogenous substances are derived from the proteins of the latex. 6. Decomposition appears to be connected with the formation of a considerable amount of acids. 7. The production of volatile nitrogen bases as a rule accompanies the decomposition processes. These volatile products have not been identified. 8. The active nitrogen bases, either already formed or derived from complex nitrogenous substances, seem to be soluble in water but only slightly soluble in acetone.


2004 ◽  
Vol 1 (1) ◽  
pp. 110-115
Author(s):  
Baghdad Science Journal

We found that 4,5- diphenyl- 3(2- propynyl) thio- 1??-triazole [1? forms a complex with Pd (11) ion of ratio 1:1 which absorbs light in CH2CI2 at 400 nm, and 4,5- diphenyl- 3(2- propenyl) thio- 1,2,4- triazole [II] forms complexes with Pd (II) ion of ratio 1:1 which absorbs light at 390 nm, and of ratio 2:1 which absorbs light at 435 nm. On the other hand, we found that the new derivative 4- phenyl- 5( p- amino phenyl) -3- mercapto- 1,2,4- triazole ?111? forms complexes with Cu (II) ion of the ratio 1:1 which absorbs light at 380 nm, with Ni (II) ion of the ratio 3:1 which absorbs light at 358 nm; and with Co (11) ion of the ratio 3.2:1 which absorbs light at 588 nm. The ratio of the complexes were determined by measuring the electronic spectra of the complexes in CH2G2 and (CH^NCHO at different concentrations ofthe ligands and f?xed ' •' of the metal ion in every case, then applying the molar ratio plots on the data. Our results were confirmed by precipitating most ofthe above complexes in solid state, and then each complex was analyzed elementally.


2011 ◽  
Vol 29 (No. 4) ◽  
pp. 373-381 ◽  
Author(s):  
J. Horníčková ◽  
R. Kubec ◽  
J. Velíšek ◽  
CejpekK ◽  
J. Ovesná ◽  
...  

The contents of three S-alk(en)ylcysteine sulfoxides (alliin, methiin, and isoalliin) were determined in the leaves, pseudostems, and bulbs of six garlic genotypes (two flowering plant morphotypes, two semi bolters, and two scape absent morphotypes) cultivated for five consecutive years at the same location. The average levels of alliin, methiin, and isoalliin were found to be as follows: 1.92, 0.44, and 0.07 mg/g fw in the leaves, 1.57, 0.27, and 0.08 mg/g fw in the pseudostems, and 1.71, 0.20 and 0.13 mg/g fw in the bulbs, respectively. No statistically significant year-to-year differences were observed between the samples. Furthermore, the total contents and relative proportions of S-alk(en)ylcysteine sulfoxides in various parts of the plants (leaves, pseudostems, bulbs and roots) were evaluated in detail during the whole vegetation period. It was observed that the total content of these amino acids gradually decreased in all parts except for the bulbs. In the bulbs, the total content initially decreased after planting but significantly increased in June and culminated before harvest. Analogous trends were also observed for alliin and methiin concentrations. On the other hand, isoalliin levels steadily decreased during the whole vegetation period in all parts of the plants.


2001 ◽  
Vol 276 (15) ◽  
pp. 11705-11711 ◽  
Author(s):  
Jan B. Koenderink ◽  
Herman G. P. Swarts ◽  
H. Christiaan Stronks ◽  
Harm P. H. Hermsen ◽  
Peter H. G. M. Willems ◽  
...  

In this study we reveal regions of Na+,K+-ATPase and H+,K+-ATPase that are involved in cation selectivity. A chimeric enzyme in which transmembrane hairpin M5-M6 of H+,K+-ATPase was replaced by that of Na+,K+-ATPase was phosphorylated in the absence of Na+and showed no K+-dependent reactions. Next, the part originating from Na+,K+-ATPase was gradually increased in the N-terminal direction. We demonstrate that chimera HN16, containing the transmembrane segments one to six and intermediate loops of Na+,K+-ATPase, harbors the amino acids responsible for Na+specificity. Compared with Na+,K+-ATPase, this chimera displayed a similar apparent Na+affinity, a lower apparent K+affinity, a higher apparent ATP affinity, and a lower apparent vanadate affinity in the ATPase reaction. This indicates that theE2K form of this chimera is less stable than that of Na+,K+-ATPase, suggesting that it, like H+,K+-ATPase, de-occludes K+ions very rapidly. Comparison of the structures of these chimeras with those of the parent enzymes suggests that the C-terminal 187 amino acids and the β-subunit are involved in K+occlusion. Accordingly, chimera HN16 is not only a chimeric enzyme in structure, but also in function. On one hand it possesses the Na+-stimulated ATPase reaction of Na+,K+-ATPase, while on the other hand it has the K+occlusion properties of H+,K+-ATPase.


2007 ◽  
Vol 277 ◽  
pp. 1-3 ◽  

In a nutshellExercise increases oxidation of amino acids such as leucine.On the other hand, leucine is a powerful mediator of protein synthesis in muscle, promoting improved function, reduced fatigue and enhanced recovery from exertion. There is RCT evidence that leucine improves sports performance, particularly for endurance activity, but more evidence is still needed.


1993 ◽  
Vol 58 (6) ◽  
pp. 1445-1451 ◽  
Author(s):  
Lubomír Lochmann ◽  
Hana Jakubův ◽  
Lambert Brandsma

The presence or absence of lithium tert-pentoxide (t-PeOLi) in Superbases, prepared from butyllithium (BuLi) and potasium tert-pentoxide (t-PeOK), affects the metallation rate of toluene only slightly, the rate factors being between 1.0 and 0.3. Thus, lithium alkoxide is not essential for the high reactivity of Superbases. However, the ring-to-side chain metallation ratio is increased in the presence of t-PeOLi. On the other hand, an excess of t-PeOK (molar ratio t-PeOK/BuLi ≥ 3)considerably increases in the rate and yield of the side chain methallation of toluene and suppresses its ring mrtallation. In this way, alkoxides enable some control on regioselectivity in Superbase reactions.


2013 ◽  
Vol 9 ◽  
pp. 832-837 ◽  
Author(s):  
Vera Lúcia Patrocinio Pereira ◽  
André Luiz da Silva Moura ◽  
Daniel Pais Pires Vieira ◽  
Leandro Lara de Carvalho ◽  
Eliz Regina Bueno Torres ◽  
...  

New chiral (S,E)-γ-N,N-dibenzylated nitroalkenes 2a–c were synthesized from natural L-(α)-amino acids in five steps with overall yields of 68–88%. The conjugate addition of hydride, methoxide, nitronate and azide nucleophiles to 2a–c led to the corresponding chiral 1,3-nitroamines in 74–90% yield. The conjugate addition of cyanide anion to 2a,b was followed by HNO2 elimination affording chiral aminated acrylonitriles (73–98%). On the other hand, the azide anion reacted with 2a, in acetonitrile, via a [3 + 2]-cycloaddition in which HNO2 was lost, providing the corresponding 1,2,3-triazole derivative. Direct reduction of 1,3-nitroamine derivatives 9a,b produced the corresponding 1,3-diamines in good yields.


1941 ◽  
Vol 14 (2) ◽  
pp. 300-307 ◽  
Author(s):  
R. F. A. Altman

Abstract In a previous paper the separation and analysis of proteins remaining in rubber serum, after coagulating the rubber with formic acid, were described. The present paper is concerned with the properties and identification of the products thus isolated. As for nonamino acids, it may be recalled that these are composed mainly of the unsaponifiable thick brownish red oil A4. Of the other byproducts remaining to be identified, which were designated as BII, BIII, BV and BVI, only small amounts were obtained. Of the amino acids, alanine, arginine, asparagic acid, glutamic acid, histidine, leucine, ornithine, hydroxyproline, proline, tyrosine and valine were isolated in weighable amounts. In addition to these, it was possible to identify dihydroxyphenylalanine. In this connection the results obtained by Midgley, Henne and Renoll are of interest. These investigators found, as the principal components of proteins occurring in crepe rubber, the following amino acids: arginine, asparagic acid, glycine, histidine, leucine, lysine and proline. They considered it highly probable that alanine, phenylalanine, hydroxyproline and serine were also present in these proteins. On the other hand, it was definitely established that cystine, glutamic acid and tyrosine were not present. Table I summarizes the results of Midgley and his coworkers in comparison with ours. From this table it is evident that certain proteins, viz., those containing, among other compounds, tyrosine, glutamic acid and valine, do not coagulate with the rubber, but remain in the serum. The absence of sulfur-containing amino acids can probably be attributed to certain shortcomings in the analytical method. At any rate, the presence of sulfur in proteins precipitated from latex was established when the test was repeated.


1970 ◽  
Vol 43 (4) ◽  
pp. 501-512 ◽  
Author(s):  
Samina Ahmed ◽  
Mainul Ahsan

Ca-hydroxyapatite (HA) was synthesised from egg shell under different conditions and characterised by using AAS, FT-IR, XRD, SEM and EDS techniques. The results of these analyses combined together indicate that sintered HA sample1 resembles the feature of pure and single apatite phase having hexagonal structure. A Ca/P molar ratio of 1.69 was achieved in this case. On the other hand HA obtained from calcined egg shell is a combination of both amorphous and crystalline phase and revealed a sub-stoichiometric Ca-deficient apatite form with a Ca/P ratio of 1.50. Keywords: Ca-hydroxyapatite, Biomaterials, Hexagonal, stoichiometric apatite, Ca-deficient apatite   doi: 10.3329/bjsir.v43i4.2240 Bangladesh J. Sci. Ind. Res. 43(4), 501-512, 2008


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