A set of physicochemical properties describing a protein of known structure is employed for a calibrative approach to protein solubility. Common hydrodynamic and electrophoretic properties routinely measured in the bio-analytical laboratory such as zeta potential, dipole moment, the second osmotic virial coefficient are first estimated in silico as a function a pH and solution ionic strength st...

Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning ...

Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the fusion proteins often re-introduces problems with solubility and stability. Here we describe the use...

Certain highly soluble proteins, such as Escherichia coli maltose-binding protein (MBP), have the ability to enhance the solubility of their fusion partners, making them attractive vehicles for the production of recombinant proteins, yet the mechanism of solubility enhancement remains poorly understood. Here, we report that the solubility-enhancing properties of MBP are dramatically affected by...

Phenolic compounds are removed from plant protein extracts because their interaction with proteins can lead to undesirable sensory and techno-functional changes. The trend toward less refined fractions requires clarification of the degree which removal is necessary. Chlorogenic acid (CGA) was added sunflower solutions obtain apparent CGA-protein molar ratios between 1:10 10:1. samples were incu...

For many years, the aggregation of proteins and polypeptides remained a neglected area of protein chemistry. It was only with the discovery that the insoluble deposits found in the organs and tissues of patients suffering from different diseases were enriched in a single, but different polypeptide, that the interest in understanding how and why these proteins aggregate arose (Fernàndez-Busquets...

One of the most vexing problems facing structural genomics efforts and the biotechnology enterprise in general is the inability to efficiently produce functional proteins due to poor folding and insolubility. Additionally, protein misfolding and aggregation has been linked to a number of human diseases, such as Alzheimer's. Thus, a robust cellular assay that allows for direct monitoring, manipu...

1. Introduction A major impediment to the production of recombinant proteins in Escheri-chia coli is their tendency to accumulate in the form of insoluble and biologically inactive aggregates known as inclusion bodies. Although it is sometimes possible to convert aggregated material into native, biologically-active protein , this is a time consuming, labor-intensive, costly, and uncertain under...

Northrop and coworkers (1-3) believe that solubility studies constitute the most sensitive and reliable test for determining the homogeneity of proteins and have used them successfully as a test for purity of their crystalline enzyme preparations. Herriott, Desreux, and Northrop (4) were able to show that a number of pepsin solutions that appeared to be homogeneous in electrophoresis experiment...

Preventing protein aggregation is a major goal of biotechnology. Since protein aggregates are mainly comprised of unfolded proteins, protecting against denaturation is likely to assist solubility in an aqueous medium. Contrary to this concept, we found denatured total cellular protein mixture from mammalian cell kept high solubility in pure water when the mixture was nucleic acids free. The lys...