Practical protocols for production of very high yields of recombinant proteins using Escherichia coli.

The gram-negative bacterium Escherichia coli offers a mean for rapid, high yield, and economical production of recombinant proteins. However, high-level production of functional eukaryotic proteins in E. coli may not be a routine matter, sometimes it is quite challenging.

An efficient on-column expressed protein ligation strategy: application to segmental triple labeling of human apolipoprotein E3.

Expressed protein ligation (EPL) is an intein-based approach that has been used for protein engineering and biophysical studies of protein structures. One major problem of the EPL is the low yield of final ligation product, primarily due to the complex procedure of the EPL, preventing EPL from gaining popularity in the research community. Here we report an efficient on-column EPL strategy, which focuses on enhancing the expression level of the intein-fusion protein that generates thioester for the EPL.

A monomeric, biologically active, full-length human apolipoprotein E.

Apolipoprotein E (apoE) is an exchangeable apolipoprotein that plays an important role in lipid/lipoprotein metabolism and cardiovascular diseases. Recent evidence indicates that apoE is also critical in several other important biological processes, including Alzheimer's disease, cognitive function, immunoregulation, cell signaling, and infectious diseases. Although the X-ray crystal structure of the apoE N-terminal domain was solved in 1991, the structural study of full-length apoE is hindered by apoE's oligomerization property.

Elimination of in vivo cleavage between target protein and intein in the intein-mediated protein purification systems.

IMPACT is a novel protein purification system developed by New England BioLabs. This system utilizes the inducible self-cleavage activity of a protein splicing element (termed intein) to separate the target protein from the affinity intein-CBD tag without using any proteases. The IMPACT system has been widely used as an easy, one-step protein purification procedure.