Structure/function studies of dogfish α-crystallin, comparison with bovine α-crystallin

Purpose: α-Crystallin is the major protein of the mammalian lens where it contributes to the refractive properties needed for vision and possibly to the stability of the tissue. The aim of this study was to determine whether the properties of α-crystallin have changed during the course of evolution. Methods: Dogfish α-crystallin, which appeared over 420 million years ago, has been contrasted with bovine α-crystallin, which emerged around 160 million years later, by comparing their sizes, the microenvironments of their cysteine and tryptophan residues, their chaperone-like activities and the flexibility of their COOH-terminal extensions. Results: Dogfish α-crystallin consists of α A- and α B-polypeptides, in a 1: 5 ratio, and has a molecular mass of around 400 kDa. By contrast, the bovine protein is around 600-800 kDa in mass and has a 3: 1 subunit ratio. Cysteine residues in the proteins were equally accessible to reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). Quenching of fluorescence with acrylamide indicated tryptophan residues in the two proteins were in similar environments. The chaperone activity of dogfish α-crystallin was comparable to that of bovine α-crystallin in preventing the heat-induced precipitation of β(L)-crystallin but the dogfish protein was three times more effective at preventing insulin precipitation after reduction at 37 degrees C. H-1 nuclear magnetic resonance spectroscopic studies showed that the last 17 amino acids of the dogfish α B polypeptide (V162-K178) have great conformational flexibility, are highly exposed to solvent and adopt little ordered conformation. This is comparable to, but slightly longer in length, than the COOH-terminal extension observed in mammalian alpha-crystallins. Conclusions: The structure and properties of α-crystallin have changed relatively little during the evolutionary period from the emergence of sharks and mammals. © US National Library of Medicine National Institutes of Healt

The effect of dextran on subunit exchange of αA-crystallin

α-Crystallin, a member of small heat shock protein (sHsp) family, is comprised of αA and αB subunits and acts as a molecular chaperone by interacting with unfolding proteins to prevent their aggregation. The αA-crystallin homopolymer consists of 30-40 subunits that are undergoing dynamic exchange. α-Crystallin and αA-crystallin are poorer chaperones in the presence of the crowding agent, dextran. Using fluorescence resonance energy transfer, it is shown that the αAcrystallin subunit exchange rate strongly increased with temperature. Binding of reduced ovotransferrin to αA-crystallin markedly decreases the rate of subunit exchange, as does the presence of dextran. In addition, in the presence of dextran the effect of reduced ovotransferrin on decreasing the rate of subunit exchange of αA-crystallin is stronger than in the absence of dextran. Under the conditions of molecular crowding, the αA-crystallin subunit exchange rate is not temperature-dependent. The exchange rate of αA-crystallin subunits correlates with its chaperone efficiency i.e. the variation in chaperone ability of αA-crystallin increases with temperature. However, in the presence of dextran the temperature dependence of the chaperone action of αA-crystallin is eliminated

Structure/function studies of dogfish alpha-crystallin, comparison with bovine alpha-crystallin

Purpose: α-Crystallin is the major protein of the mammalian lens where it contributes to the refractive properties needed for vision and possibly to the stability of the tissue. The aim of this study was to determine whether the properties of α-crystallin have changed during the course of evolution. Methods: Dogfish α-crystallin, which appeared over 420 million years ago, has been contrasted with bovine α-crystallin, which emerged around 160 million years later, by comparing their sizes, the microenvironments of their cysteine and tryptophan residues, their chaperone-like activities and the flexibility of their COOH-terminal extensions. Results: Dogfish α-crystallin consists of αA- and αB-polypeptides, in a 1:5 ratio, and has a molecular mass of around 400 kDa. By contrast, the bovine protein is around 600-800 kDa in mass and has a 3:1 subunit ratio. Cysteine residues in the proteins were equally accessible to reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). Quenching of fluorescence with acrylamide indicated tryptophan residues in the two proteins were in similar environments. The chaperone activity of dogfish α-crystallin was comparable to that of bovine α-crystallin in preventing the heat-induced precipitation of βL-crystallin but the dogfish protein was three times more effective at preventing insulin precipitation after reduction at 37 ˚C. 1H nuclear magnetic resonance spectroscopic studies showed that the last 17 amino acids of the dogfish αB polypeptide (V162-K178) have great conformational flexibility, are highly exposed to solvent and adopt little ordered conformation. This is comparable to, but slightly longer in length, than the COOH-terminal extension observed in mammalian α-crystallins. Conclusions: The structure and properties of α-crystallin have changed relatively little during the evolutionary period from the emergence of sharks and mammals.A. Ghahghaei, A. Rekas, J.A. Carver, R.C. Augustey

Inhibitory effect of crocin(s) on lens crystallin glycation and aggregation, results in the decrease of the risk of diabetic cataract

The current study investigates the inhibitory effect of crocin(s), also known as saffron apocarotenoids, on protein glycation and aggregation in diabetic rats, and α-crystallin glycation. Thus, crocin(s) were administered by intraperitoneal injection to normal and streptozotocin-induced diabetic rats. The cataract progression was recorded regularly every two weeks and was classified into four stages. After eight weeks, the animals were sacrificed and the parameters involved in the cataract formation were measured in the animal lenses. Some parameters were also determined in the serum and blood of the rats. In addition, the effect of crocin(s) on the structure and chaperone activity of α-crystallin in the presence of glucose was studied by different methods. Crocin(s) lowered serum glucose levels of diabetic rats and effectively maintained plasma total antioxidants, glutathione levels and catalase activity in the lens of the animals. In the in vitro study, crocin(s) inhibited α-crystallin glycation and aggregation. Advanced glycation end products fluorescence, hydrophobicity and protein cross-links were also decreased in the presence of crocin(s). In addition, the decreased chaperone activity of α-crystallin in the presence of glucose changed and became close to the native value by the addition of crocin(s) in the medium. Crocin(s) thus showed a powerful inhibitory effect on α-crystallin glycation and preserved the structure-function of this protein. Crocin(s) also showed the beneficial effects on prevention of diabetic cataract. © 2016 by the authors

The Comparison of Yield and Yield Components of Lentil Genotypes at Low Irrigation in Sistan Region

Abstract\ud The important factors for high seed yielding genotypes selection, involve of those characteristics that affecting on seed per plants. In order to evaluation and selection the fittest lentil genotypes in sistan region, an experiment with 10 genotyps were carried out in agricultural research farm at Agricultural research Institute of Zabol University, in 2006-2007 by complete randomize blocks design in three replications.The results showed significantly differences (

Optimal (R, Q) policy and pricing for two-echelon supply chain with lead time and retailer’s service-level incomplete information

Abstract Many studies focus on inventory systems to analyze different real-world situations. This paper considers a two-echelon supply chain that includes one warehouse and one retailer with stochastic demand and an up-to-level policy. The retailer’s lead time includes the transportation time from the warehouse to the retailer that is unknown to the retailer. On the other hand, the warehouse is unaware of retailer’s service level. The relationship between the retailer and the warehouse is modeled based on the Stackelberg game with incomplete information. Moreover, their relationship is presented when the warehouse and the retailer reveal their private information using the incentive strategies. The optimal inventory and pricing policies are obtained using an algorithm based on bi-level programming. Numerical examples, including sensitivity analysis of some key parameters, will compare the results between the Stackelberg models. The results show that information sharing is more beneficial to the warehouse rather than the retailer