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Ca-induced renaturation of Bacillus licheniformis alpha-amylase in the presence of urea has been employed to determine the binding constants of the ion. The native enzyme is folded at 3M urea while the Ca-depleted protein is largely unfolded at this denaturant concentration. Refolding of the protein has been monitored by circular dichroism and the titration curves have been analyzed assuming a model of three independent binding sites. The stoichiometry has been taken from X-ray studies. The refolded protein exhibits a secondary structure that is similar but not identical to that of the native protein. The binding constants have been used to construct a phase diagram that illustrates the contribution of Ca-binding to the resistance against urea unfolding. 相似文献
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Fernando López-Soto Nidia León-Sicairos Kamran Nazmi Jan G. Bolscher Mireya de la Garza 《Biometals》2010,23(3):563-568
Entamoeba histolytica is a parasitic protozoan that produces amoebiasis, an intestinal disease characterized by ulcerative colitis and dysentery.
In some cases, trophozoites can travel to the liver leading to hepatic abscesses and death. Recently, lactoferrin and lactoferricin
B have been shown to be amoebicidal in axenic cultures. The aim of this work was to determine whether the lactoferrin-peptides
lactoferricin amino acids 17–30, lactoferrampin amino acids 265–284, and lactoferrin chimera which is a fusion product of
the two peptides, are capable of producing a microbicidal effect to trophozoites of E. histolytica. We evaluated the killing effect of these peptides in growth kinetics carried out in axenic culture medium to which different
concentrations of peptides were added. At 50 μM of peptide concentration, lactoferricin and lactoferrampin had a moderate
amoebicidal effect, since a 45–50% of trophozoites remained viable at 24 h culture. However, at 50 μM of the lactoferrin chimera
75% amoeba were killed whereas at 100 μM all cells died. These data indicate that of lactoferrin-peptides mainly the chimera
have amoebicidal activity in a time- and concentration-dependent manner. The lactoferrin-peptides might be useful as therapeutic
agents against amoebiasis and thereby diminish the use of metronidazole, which is extremely toxic for the host. 相似文献
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Ahmad Nazmi Rosli Maizathul Akmam Abu Bakar Vannajan Sanghiran Lee Sharifuddin Md Zain Mohd Rais Ahmad Ninie Suhana Abdul Manan Yatimah Alias Pei Meng Woi 《Journal of molecular modeling》2014,20(9):1-13
Amino acids are constituents of proteins and enzymes which take part almost in all metabolic reactions. Glutamic acid, with an ability to form a negatively charged side chain, plays a major role in intra and intermolecular interactions of proteins, peptides, and enzymes. An exhaustive conformational analysis has been performed for all eight possible forms at B3LYP/cc-pVTZ level. All possible neutral, zwitterionic, protonated, and deprotonated forms of glutamic acid structures have been investigated in solution by using polarizable continuum model mimicking water as the solvent. Nine families based on the dihedral angles have been classified for eight glutamic acid forms. The electrostatic effects included in the solvent model usually stabilize the charged forms more. However, the stability of the zwitterionic form has been underestimated due to the lack of hydrogen bonding between the solute and solvent; therefore, it is observed that compact neutral glutamic acid structures are more stable in solution than they are in vacuum. Our calculations have shown that among all eight possible forms, some are not stable in solution and are immediately converted to other more stable forms. Comparison of isoelectronic glutamic acid forms indicated that one of the structures among possible zwitterionic and anionic forms may dominate over the other possible forms. Additional investigations using explicit solvent models are necessary to determine the stability of charged forms of glutamic acid in solution as our results clearly indicate that hydrogen bonding and its type have a major role in the structure and energy of conformers. 相似文献