SUMO fusion technology for difficult-to-express proteins

Abstract

The demands of structural and functional genomics for large quantities of soluble, properly folded proteins in heterologous hosts have been aided by advancements in the field of protein production and purification. Escherichia coli, the preferred host for recombinant protein expression, presents many challenges which must be surmounted in order to over-express heterologous proteins. These challenges include the proteolytic degradation of target proteins, protein misfolding, poor solubility, and the necessity for good purification methodologies. Gene fusion technologies have been able to improve heterologous expression by overcoming many of these challenges. The ability of gene fusions to improve expression, solubility, purification, and decrease proteolytic degradation will be discussed in this review. The main disadvantage, cleaving the protein fusion, will also be addressed. Focus will be given to the newly described SUMO fusion system and the improvements that this technology has advanced over traditional gene fusion systems.

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

“…When used as an N-terminal carrier protein during prokaryotic expression, SUMO promotes folding and structural stability, which leads to enhanced functional production compared to untagged protein [48][49][50][51][52][53][54]. Unlike GST and MBP, SUMO does not itself serve as a means for purification of fusion proteins; however, the His 6 in series with the SUMO tag has been established to facilitate purification of fusion proteins.…”

Soluble expression and purification of heparinase I in Escherichia coli using a hexahistidine-tagged small ubiquitin-like modifier as a fusion partner

“…Soluble partners are able to enhance the solubility of recombinant protein, such as glutathione S-transferase (GST), maltose-binding protein (MBP), thioredoxin (Trx), etc [12,13]. Among the expression partners, SUMO, a small ubiquitin-modifying protein which can dramatically improve the soluble expression, has attracted a lot of attention recently [14,15]. A comparison between SUMO fusion technologies and traditional gene fusion systems implies that SUMO is superior to traditional fusion tags in enhancing protein expression and solubility [14].…”

An Amyloidogenic Sequence at the N-Terminus of the Androgen Receptor Impacts Polyglutamine Aggregation

“…These experiments, conducted with peptides of various lengths, identified the sequence KELCKAVSVSM as the minimal motif for the formation of amyloid fibers upon addition of 10% DMSO. To determine whether the amyloidogenic property of this sequence is retained in a larger peptide, in the context of a fusion protein, we fused the peptide to a SUMO protein and expressed it in E coli [21]. After affinity purification of the bacterial cell lysates on nickel agarose resins, a major product with the expected mass of the HisSUMO-peptide fusion protein of 15.819 kDa was identified on a SDS-PAGE gel ( Figure 1A, lane 5).…”