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A double-headed chymotrypsin inhibitor, WCI, from winged bean seeds was cloned for structural and biochemical studies. The inhibitor was subjected to two point mutations at a conserved position, Asn14. This residue, known to have a pivotal role in stabilizing the first reactive-site loop...
The Asn108β→Lys mutation in hemoglobin (HbPresbyterian mutation) endows a low O 2 affinity-inducing propensity to the protein. Introduction of a fumaryl cross-bridge between its two α99 lysine residues also induces a low O 2 affinity into HbA. We have now engineered an αα-fumaryl...
Solvent entropy is a force to consider in protein folding and protein design but is difficult to model. It is investigated here in the context of the hp model: Two types of residues, hydrophobic and hydrophilic, are modeled on a lattice. Nine chains and two- and three-dimensional simulations are...
We examined several strategies for the secretion of Kluyveromyces lactis β-galactosidase into the culture medium, in order to facilitate the downstream processing and purification of this intracellular enzyme of great industrial interest. We constructed plasmids by fusing the LAC4 gene or...
The biological activity of some proteins is known to be sensitive to oxidative damage caused by a variety of oxidants. The model protein staphylococcal nuclease was used to explore the effect on protein structural stability of oxidizing methionine to the sulfoxide form. These effects were...
The `FLITRX' random peptide library, consisting of dodecamer loop peptides displayed on a thioredoxin-flagellin scaffold on Escherichia coli , was used to select peptide sequences with affinity for a monoclonal antibody. These peptides were further screened for pH- and metal-sensitive antibody...
The observation that strong amphiphilic α-helical potential exists in all proteins, including β-sheet proteins, has given rise to the idea that α-helical intermediates may be critical to the folding paths of all proteins. Here we report that regions with amphiphilic α-helical potential in...
The distribution of the χ 1 , χ 2 dihedral angles in a dataset consisting of 12 unrelated 4-α-helical bundle proteins was determined and qualitatively compared with that observed in globular proteins. The analysis suggests that the 4-α-helical bundle motif could occasionally impose steric...
Endothelin-converting enzyme 1 (ECE-1, EC 22.214.171.124) is a zinc-dependent type II mammalian membrane protein comprising the active site in the ectodomain. It exists in multiple splice variants that all catalyze the last and rate-limiting step in the activation of preproendothelin to the highly...
Three-phase partitioning is fast developing as a novel bioseparation strategy with a wide range of applications including enzyme stability and enhancement of its catalytic activity. Despite all this, the enzyme behaviour in this process still remains unknown. A serine proteinase, proteinase K,...
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