Understanding the connection between local and global dynamics can provide valuable insights into enzymatic function may contribute to development of novel strategies for enzyme modulation. In this work, we investigated at both (active site) levels Shikimate Kinase (SK) through microsecond time-scale molecular (MD) simulations holoenzyme in product state. Our focus was on wild-type (WT) two mut...

Genetic studies of bacteria and eukaryotes have led to identification of several gene products that are involved in the biosynthesis of protein-bound iron-sulfur clusters. One of these proteins, ISU, is homologous to the N-terminus of bacterial NifU. The mature forms of His-tagged wild-type and D37A Schizosaccharomyces pombe ISU1 were cloned and overexpressed as inclusion bodies in Escherichia ...

We study apo and holo forms of the bacterial ferric binding protein (FBP) which exhibits the so-called ferric transport dilemma: it uptakes iron from the host with remarkable affinity, yet releases it with ease in the cytoplasm for subsequent use. The observations fit the "conformational selection" model whereby the existence of a weakly populated, higher energy conformation that is stabilized ...

BACKGROUND Measurement of serum cobalamin levels is routinely used to diagnose cobalamin deficiency. Surprisingly, approximately 15% of patients have high cobalamin levels and no consensus exists regarding the clinical implications. METHODS Hospital-treated patients above 18 years of age referred for serum cobalamin measurement were included in groups of patients [percentage cobalamin supplem...

Intrinsically disordered proteins do not adopt well-defined native structures and therefore present an intriguing challenge in terms of structural elucidation as they are relatively inaccessible to traditional approaches such as NMR and X-ray crystallography. Many members of this important group of proteins have a distinct biological function and frequently undergo a conformational change on bi...

Apo-Calmodulin acts as the light chain for unconventional myosin V, and treatment with Ca(2+) can cause dissociation of calmodulin from the 6IQ region of the myosin heavy chain. The effects of Ca(2+) on the stoichiometry and affinity of interactions of calmodulin and its two domains with two myosin-V peptides (IQ3 and IQ4) have therefore been quantified in vitro, using fluorescence and near- an...

The objective of this study was to develop a simple and economical protocol for separating ovotransferrin from egg white. Egg white was separated from the yolk and diluted with the same volume of distilled water. To prevent denaturation during the separation process, ovotransferrin in 2x-diluted egg white was converted to its holo-form by adding 20 mM FeCl3.6H2O solution (0.25 to 3 mL/100 mL). ...

To determine the nature of binding of transcobalamin II (TC-II) to liver cells, we covalently coupled purified holo-TC-II to submicron latex minibeads using glutaraldehyde. Incubation of the probe with liver cell suspensions at 4 degrees C led to its binding by endothelial cells but not by hepatocytes or Kupffer cells, as visualized by scanning electron microscopy. At 37 degrees C, the probe wa...